CN101218256B - Antibodies against cd38 for treatment of multiple myeloma - Google Patents

附图简要说明Brief description of the drawings

图1A所示的是通过流式细胞仪测定的-003、-005和同型体对照抗体HuMab-KLH与CD38转染的CHO(CHO-CD38)细胞的结合。实验设置在实施例4中描述。Figure 1A shows the binding of -003, -005 and the isotype control antibody HuMab-KLH to CD38 transfected CHO (CHO-CD38) cells measured by flow cytometry. The experimental setup is described in Example 4.

图1B所示的是通过流式细胞仪测定的-024和HuMab-KLH与CD38转染的CHO(CHO-CD38)细胞的结合。实验设置在实施例4中描述。Figure 1B shows the binding of -024 and HuMab-KLH to CD38 transfected CHO (CHO-CD38) cells measured by flow cytometry. The experimental setup is described in Example 4.

图2A所示的是通过流式细胞仪测定的-003、-005和同型体对照抗体HuMab-KLH与Daudi细胞的结合。实验设置在实施例4中描述。Figure 2A shows the binding of -003, -005 and the isotype control antibody HuMab-KLH to Daudi cells as determined by flow cytometry. The experimental setup is described in Example 4.

图2B所示的是通过流式细胞仪测定的-024和HuMab-KLH与Daudi细胞的结合。实验设置在实施例4中描述。Figure 2B shows the binding of -024 and HuMab-KLH to Daudi cells measured by flow cytometry. The experimental setup is described in Example 4.

图3所示的是-003、-005、-024和HuMab-KLH与多发性骨髓瘤细胞的结合。实验设置在实施例4中描述。Figure 3 shows the binding of -003, -005, -024 and HuMab-KLH to multiple myeloma cells. The experimental setup is described in Example 4.

图4A所示的是与利妥昔单抗和HuMab-KLH相比较，-003和-005通过ADCC诱导Daudi细胞裂解的能力。实验设置在实施例5中描述。Figure 4A shows the ability of -003 and -005 to induce Daudi cell lysis via ADCC compared to rituximab and HuMab-KLH. The experimental setup is described in Example 5.

图4B所示的是与HuMab-KLH相比较，-024通过ADCC诱导Daudi细胞裂解的能力。实验设置在实施例5中描述。Figure 4B shows the ability of -024 to induce Daudi cell lysis via ADCC compared to HuMab-KLH. The experimental setup is described in Example 5.

图5A所示的是与HuMab-KLH相比较，-003、-005和-024通过ADCC诱导新鲜的多发性骨髓瘤细胞裂解的能力。实验设置在实施例5中描 述。Figure 5A shows the ability of -003, -005 and -024 to induce lysis of fresh multiple myeloma cells by ADCC compared to HuMab-KLH. The experimental setup is described in Example 5.

图5B所示的是与HuMab-KLH相比较，-003、-005和-024通过ADCC诱导新鲜的浆细胞白血病肿瘤细胞裂解的能力。实验设置在实施例5中描述。Figure 5B shows the ability of -003, -005 and -024 to induce lysis of fresh plasma cell leukemia tumor cells by ADCC compared to HuMab-KLH. The experimental setup is described in Example 5.

图6所示的是与HuMab-KLH相比较，-003和-005通过ADCC诱导JK6L(一种多发性骨髓瘤细胞系)裂解的能力。实验设置在实施例5中描述。Figure 6 shows the ability of -003 and -005 to induce lysis of JK6L (a multiple myeloma cell line) by ADCC compared to HuMab-KLH. The experimental setup is described in Example 5.

图7所示的是与HuMab-KLH相比较，-003和-005通过ADCC诱导AMO-1(一种多发性骨髓瘤细胞系)裂解的能力。实验设置在实施例5中描述。Figure 7 shows the ability of -003 and -005 to induce lysis of AMO-1 (a multiple myeloma cell line) by ADCC compared to HuMab-KLH. The experimental setup is described in Example 5.

图8所示的是与HuMab-KLH相比较，通过-003和-005诱导的对Daudi-luc细胞的CDC介导的裂解。实验设置在实施例6中描述。Figure 8 shows CDC-mediated lysis of Daudi-luc cells induced by -003 and -005 compared to HuMab-KLH. The experimental setup is described in Example 6.

图9A所示的是与HuMab-KLH相比较，通过-003和-005诱导的对CHO-CD3 8细胞的CDC介导的裂解。实验设置在实施例6中描述。Figure 9A shows CDC-mediated lysis of CHO-CD38 cells induced by -003 and -005 compared to HuMab-KLH. The experimental setup is described in Example 6.

图9B所示的是与HuMab-KLH相比较，通过-024诱导的对CHO-CD3 8细胞的CDC介导的裂解。实验设置在实施例6中描述。Figure 9B shows CDC-mediated lysis of CHO-CD38 cells induced by -024 compared to HuMab-KLH. The experimental setup is described in Example 6.

图10A所示的是在存在-003、-005和HuMab-KLH时，3％耐受性肿瘤细胞的CDC介导的裂解。实验设置在实施例6中描述。Figure 10A shows the CDC-mediated lysis of 3% resistant tumor cells in the presence of -003, -005 and HuMab-KLH. The experimental setup is described in Example 6.

图10B所示的是在存在-003、-005和HuMab-KLH时，9％耐受性肿瘤细胞的CDC介导的裂解。实验设置在实施例6中描述。Figure 10B shows the CDC-mediated lysis of 9% of resistant tumor cells in the presence of -003, -005 and HuMab-KLH. The experimental setup is described in Example 6.

图10C所示的是在存在-003、-005和HuMab-KLH时，30-40％耐受性肿瘤细胞的CDC介导的裂解。实验设置在实施例6中描述。Figure 10C shows the CDC-mediated lysis of 30-40% of resistant tumor cells in the presence of -003, -005 and HuMab-KLH. The experimental setup is described in Example 6.

图10D所示的是在存在-003、-005和HuMab-KLH时，70％耐受性肿瘤细胞的CDC介导的裂解。实验设置在实施例6中描述。Figure 10D shows the CDC-mediated lysis of 70% of resistant tumor cells in the presence of -003, -005 and HuMab-KLH. The experimental setup is described in Example 6.

图10E所示的是在存在-024和HuMab-KLH时，多发性骨髓瘤细胞的CDC介导的裂解。实验设置在实施例6中描述。Figure 10E shows CDC-mediated lysis of multiple myeloma cells in the presence of -024 and HuMab-KLH. The experimental setup is described in Example 6.

图11所示的是-003和-005不交叉抑制对CD38的结合。实验设置在实施例7中描述。Figure 11 shows that -003 and -005 do not cross-inhibit CD38 binding. The experimental setup is described in Example 7.

图12A所示的是含-003的巨噬细胞、淋巴细胞和浆B细胞的免疫组化染色。实验设置在实施例10中描述。Figure 12A shows the immunohistochemical staining of -003-containing macrophages, lymphocytes and plasma B cells. The experimental setup is described in Example 10.

图12B所示的是含-003的支气管上皮细胞的免疫组化染色。实验设置在实施例10中描述。Figure 12B shows the immunohistochemical staining of bronchial epithelial cells containing -003. The experimental setup is described in Example 10.

图12C所示的是含-003的肌细胞的免疫组化染色。实验设置在实施例10中描述。Figure 12C shows the immunohistochemical staining of -003-containing myocytes. The experimental setup is described in Example 10.

图12D所示的是含-003的猕猴淋巴组织的免疫组化染色。实验设置在实施例10中描述。Figure 12D shows the immunohistochemical staining of -003-containing macaque lymphoid tissue. The experimental setup is described in Example 10.

图13A所示的是含-005的巨噬细胞、淋巴细胞和浆B细胞的免疫组化染色。实验设置在实施例10中描述。Figure 13A shows the immunohistochemical staining of macrophages, lymphocytes and plasma B cells containing -005. The experimental setup is described in Example 10.

图13B所示的是含-005的支气管上皮细胞的免疫组化染色。实验设置在实施例10中描述。Figure 13B shows the immunohistochemical staining of bronchial epithelial cells containing -005. The experimental setup is described in Example 10.

图13C所示的是含-005的肌细胞的免疫组化染色。实验设置在实施例10中描述。Figure 13C shows the immunohistochemical staining of -005-containing myocytes. The experimental setup is described in Example 10.

图13D所示的是含-005的猕猴淋巴组织的免疫组化染色。实验设置在实施例10中描述。Figure 13D shows the immunohistochemical staining of -005-containing macaque lymphoid tissue. The experimental setup is described in Example 10.

图14A所示的是含CD31的肝内皮细胞的免疫组化染色。实验设置在实施例10中描述。Figure 14A shows immunohistochemical staining of CD31-containing liver endothelial cells. The experimental setup is described in Example 10.

图14B所示的是含vWF的肝内皮细胞的免疫组化染色。实验设置在实施例10中描述。Figure 14B shows the immunohistochemical staining of liver endothelial cells containing vWF. The experimental setup is described in Example 10.

图14C所示的是含抗KLH的肝内皮细胞的免疫组化染色。实验设置在实施例10中描述。Figure 14C shows immunohistochemical staining of hepatic endothelial cells with anti-KLH. The experimental setup is described in Example 10.

图14D所示的是含-003的肝内皮细胞的免疫组化染色。实验设置在实施例10中描述。Figure 14D shows the immunohistochemical staining of -003-containing liver endothelial cells. The experimental setup is described in Example 10.

图14E所示的是含-005的肝内皮细胞的免疫组化染色。实验设置在实施例10中描述。Figure 14E shows the immunohistochemical staining of -005-containing liver endothelial cells. The experimental setup is described in Example 10.

图15A所示的是通过流式细胞仪测定的，与HuMab-KLH相比较，-003和-005对猕猴淋巴细胞的交叉反应性。实验设置在实施例11中描述。Figure 15A shows the cross-reactivity of -003 and -005 to macaque lymphocytes compared with HuMab-KLH as determined by flow cytometry. The experimental setup is described in Example 11.

图15B所示的是通过流式细胞仪测定的，与HuMab-KLH相比较，-003和-005对猕猴单核细胞的交叉反应性。实验设置在实施例11中描述。Figure 15B shows the cross-reactivity of -003 and -005 to macaque monocytes compared to HuMab-KLH as determined by flow cytometry. The experimental setup is described in Example 11.

图15C所示的是通过流式细胞仪测定的，与HuMab-KLH相比较，-003和-005对恒河猴PBMCs的交叉反应性。实验设置在实施例11中描述。Figure 15C shows the cross-reactivity of -003 and -005 to rhesus monkey PBMCs compared with HuMab-KLH as determined by flow cytometry. The experimental setup is described in Example 11.

图16A所示的是通过EtBr淬灭测定的-003的内在化。实验设置在 实施例12中描述。Figure 16A shows the internalization of -003 as measured by EtBr quenching. The experimental setup is described in Example 12.

图16B所示的是通过EtBr淬灭测定的-005的内在化。实验设置在实施例12中描述。Figure 16B shows the internalization of -005 as measured by EtBr quenching. The experimental setup is described in Example 12.

图17A所示的是通过体内SCID荧光成像测定的，与抗CD20单克隆抗体(利妥昔单抗)和HuMab-KLH相比较，-003和-005在预防性设置中对肿瘤细胞生长的抑制作用。实验设置在实施例13中描述。Figure 17A shows the inhibition of tumor cell growth by -003 and -005 in a prophylactic setting, as determined by in vivo SCID fluorescence imaging, compared with an anti-CD20 monoclonal antibody (rituximab) and HuMab-KLH effect. The experimental setup is described in Example 13.

图17B所示的是通过体内SCID荧光成像测定的，与抗CD20单克隆抗体(利妥昔单抗)和HuMab-KLH相比较，-003和-005在治疗性设置I中对肿瘤细胞生长的抑制作用。实验设置在实施例13中描述。Figure 17B shows the effect of -003 and -005 on tumor cell growth in therapeutic setting I compared to anti-CD20 monoclonal antibody (rituximab) and HuMab-KLH, as determined by in vivo SCID fluorescence imaging. inhibition. The experimental setup is described in Example 13.

图17C所示的是通过体内SCID荧光成像测定的，与抗CD20单克隆抗体(利妥昔单抗)和HuMab-KLH相比较，-003和-005在治疗性设置II中对肿瘤细胞生长的抑制作用。实验设置在实施例13中描述。Figure 17C shows the effect of -003 and -005 on tumor cell growth in therapeutic setting II compared to anti-CD20 mAb (rituximab) and HuMab-KLH, as determined by in vivo SCID fluorescence imaging. inhibition. The experimental setup is described in Example 13.

图17D所示的是通过体内SCID荧光成像测定的，与HuMab-KLH相比较，-003和-024在治疗性设置III中对肿瘤细胞生长的抑制作用。实验设置在实施例13中描述。Figure 17D shows the inhibitory effect of -003 and -024 on tumor cell growth in therapeutic setting III compared to HuMab-KLH as determined by in vivo SCID fluorescence imaging. The experimental setup is described in Example 13.

图18所示的是与抗CD20单克隆抗体(利妥昔单抗)和HuMab-KLH相比较，-003和-024在未交联或交联时对凋亡的诱导作用。实验设置在实施例14中描述。Figure 18 shows the induction of apoptosis by -003 and -024 when uncrosslinked or crosslinked compared with anti-CD20 monoclonal antibody (rituximab) and HuMab-KLH. The experimental setup is described in Example 14.

图19所示的是在用抗KLH(HuMab-KLH)或-005治疗14天后，在移植的RA-SCID鼠的异种移植物中CD38阳性细胞的组织学分值。方法在实施例15中描述。Figure 19 shows the histological scores of CD38 positive cells in xenografts of transplanted RA-SCID mice after 14 days of treatment with anti-KLH (HuMab-KLH) or -005. The method is described in Example 15.

图20所示的是在用抗KLH或-005治疗14天后，在移植的RA-SCID鼠的异种移植物中CD38阳性细胞的组织学分值。方法在实施例15中描述。Figure 20 shows the histological scores of CD38 positive cells in xenografts of transplanted RA-SCID mice after 14 days of treatment with anti-KLH or -005. The method is described in Example 15.

图21所示的是移植前(A)、或用抗KLH(B)或-005(C)治疗后异种移植物的B细胞CD38染色。方法在实施例15中描述。Figure 21 shows B cell CD38 staining of xenografts before transplantation (A), or after treatment with anti-KLH (B) or -005 (C). The method is described in Example 15.

图22所示的是移植前(A)、或用抗KLH(B)或-005(C)治疗后异种移植物的B细胞CD138染色。方法在实施例15中描述。Figure 22 shows B cell CD138 staining of xenografts before transplantation (A), or after treatment with anti-KLH (B) or -005 (C). The method is described in Example 15.

图23所示的是通过ELISA测定的-003和-005与野生型和突变型人CD38的结合。Figure 23 shows the binding of -003 and -005 to wild-type and mutant human CD38 as determined by ELISA.

23A：-003和-005与T237A突变型人CD38的结合。23A: Binding of -003 and -005 to T237A mutant human CD38.

23B：-003和-005与Q272R突变型人CD38的结合。23B: Binding of -003 and -005 to Q272R mutant human CD38.

23C：-003和-005与S274F突变型人CD38的结合。方法在实施例17中描述。23C: Binding of -003 and -005 to S274F mutant human CD38. The method is described in Example 17.

图24所示的是与HuMab-KLH相比较，-003和-005对人PBMCs的增殖(A)、IL-6产生(B)和IFN-γ产生(C)的作用。方法分别在实施例18、19和20中描述。Figure 24 shows the effects of -003 and -005 on the proliferation (A), IL-6 production (B) and IFN-γ production (C) of human PBMCs compared with HuMab-KLH. Methods are described in Examples 18, 19 and 20, respectively.

图25所示的是在存在各种浓度的-003(B)、-005(C)、-024(D)或抗KLH(A)时，cGDP核糖的酶产量。方法在实施例23中描述。Figure 25 shows the enzymatic production of cGDP ribose in the presence of various concentrations of -003 (B), -005 (C), -024 (D) or anti-KLH (A). The method is described in Example 23.

图26所示的是在CHO-CD38细胞(26A)的CDC、Daudi细胞的CDC(26B)、和Daudi细胞的ADCC(26C)中，-003、-005和Morphosys抗体TH-3079的比较。Figure 26 shows the comparison of -003, -005 and Morphosys antibody TH-3079 in CDC of CHO-CD38 cells (26A), CDC of Daudi cells (26B), and ADCC of Daudi cells (26C).

本发明中的序列表示在附加的序列表中。The sequences of the present invention are shown in the appended Sequence Listing.

SEQ ID No：1是抗体-003 V_L区的核苷酸序列。SEQ ID No: 1 is the nucleotide sequence of the V _L region of antibody-003.

SEQ ID No：2是抗体-003 V_L区的氨基酸序列。SEQ ID No: 2 is the amino acid sequence of the _VL region of Antibody-003.

SEQ ID No：3是抗体-003 V_L CDR1的氨基酸序列，包括SEQ IDNo：2的24-34位氨基酸。SEQ ID No: 3 is the amino acid sequence of antibody-003 V _L CDR1, including amino acids 24-34 of SEQ ID No: 2.

SEQ ID No：4是抗体-003 V_L CDR2的氨基酸序列，包括SEQ IDNo：2的50-56位氨基酸。SEQ ID No: 4 is the amino acid sequence of antibody-003 V _L CDR2, including amino acids 50-56 of SEQ ID No: 2.

SEQ ID No：5是抗体-003 V_L CDR3的氨基酸序列，包括SEQ IDNo：2的89-97位氨基酸。SEQ ID No: 5 is the amino acid sequence of antibody-003 V _L CDR3, including amino acids 89-97 of SEQ ID No: 2.

SEQ ID No：6是抗体-003 V_H区的核苷酸序列。SEQ ID No: 6 is the nucleotide sequence of the _VH region of Antibody-003.

SEQ ID No：7是抗体-003 V_H区的氨基酸序列。SEQ ID No: 7 is the amino acid sequence of the _VH region of Antibody-003.

SEQ ID No：8是抗体-003 V_H CDR1的氨基酸序列，包括SEQ IDNo：7的31-35位氨基酸。SEQ ID No: 8 is the amino acid sequence of antibody-003 V _H CDR1, including amino acids 31-35 of SEQ ID No: 7.

SEQ ID No：9是抗体-003 V_H CDR2的氨基酸序列，包括SEQ IDNo：7的50-66位氨基酸。SEQ ID No: 9 is the amino acid sequence of antibody-003 V _H CDR2, including amino acids 50-66 of SEQ ID No: 7.

SEQ ID No：10是抗体-003 V_H CDR3的氨基酸序列，包括SEQ IDNo：7的99-109位氨基酸。SEQ ID No: 10 is the amino acid sequence of antibody-003 V _H CDR3, including amino acids 99-109 of SEQ ID No: 7.

SEQ ID No：11是抗体-005 V_L区的核苷酸序列。SEQ ID No: 11 is the nucleotide sequence of the _VL region of Antibody-005.

SEQ ID No：12是抗体-005 V_L区的氨基酸序列。SEQ ID No: 12 is the amino acid sequence of the _VL region of Antibody-005.

SEQ ID No：13是抗体-005 V_L CDR1的氨基酸序列，包括SEQ IDNo：12的24-34位氨基酸。SEQ ID No: 13 is the amino acid sequence of antibody-005 V _L CDR1, including amino acids 24-34 of SEQ ID No: 12.

SEQ ID No：14是抗体-005 V_L CDR2的氨基酸序列，包括SEQ ID No：12的50-56位氨基酸。SEQ ID No: 14 is the amino acid sequence of antibody-005 V _L CDR2, including amino acids 50-56 of SEQ ID No: 12.

SEQ ID No：15是抗体-005 V_L CDR3的氨基酸序列，包括SEQ IDNo：12的89-97位氨基酸。SEQ ID No: 15 is the amino acid sequence of antibody-005 V _L CDR3, including amino acids 89-97 of SEQ ID No: 12.

SEQ ID No：16是抗体-005 V_H区的核苷酸序列。SEQ ID No: 16 is the nucleotide sequence of the _VH region of Antibody-005.

SEQ ID No：17是抗体-005 V_H区的氨基酸序列。SEQ ID No: 17 is the amino acid sequence of the _VH region of Antibody-005.

SEQ ID No：18是抗体-005 V_H CDR1的氨基酸序列，包括SEQ IDNo：17的31-35位氨基酸。SEQ ID No: 18 is the amino acid sequence of antibody-005 V _H CDR1, including amino acids 31-35 of SEQ ID No: 17.

SEQ ID No：19是抗体-005 V_H CDR2的氨基酸序列，包括SEQ IDNo：17的50-66位氨基酸。SEQ ID No: 19 is the amino acid sequence of antibody-005 V _H CDR2, including amino acids 50-66 of SEQ ID No: 17.

SEQ ID No：20是抗体-005 V_H CDR3的氨基酸序列，包括SEQ IDNo：17的99-111位氨基酸。SEQ ID No: 20 is the amino acid sequence of antibody-005 V _H CDR3, including amino acids 99-111 of SEQ ID No: 17.

SEQ ID No：21是抗体-024 V_L区的核苷酸序列。SEQ ID No: 21 is the nucleotide sequence of the _VL region of Antibody-024.

SEQ ID No：22是抗体-024 V_L区的氨基酸序列。SEQ ID No: 22 is the amino acid sequence of the _VL region of Antibody-024.

SEQ ID No：23是抗体-024 V_L CDR1的氨基酸序列，包括SEQ IDNo：22的24-34位氨基酸。SEQ ID No: 23 is the amino acid sequence of Antibody-024 V _L CDR1, including amino acids 24-34 of SEQ ID No: 22.

SEQ ID No：24是抗体-024 V_L CDR2的氨基酸序列，包括SEQ IDNo：22的50-66位氨基酸。SEQ ID No: 24 is the amino acid sequence of antibody-024 V _L CDR2, including amino acids 50-66 of SEQ ID No: 22.

SEQ ID No：25是抗体-024 V_L CDR3的氨基酸序列，包括SEQ IDNo：22的89-97位氨基酸。SEQ ID No: 25 is the amino acid sequence of antibody-024 V _L CDR3, including amino acids 89-97 of SEQ ID No: 22.

SEQ ID No：26是抗体-024 V_H区的核苷酸序列。SEQ ID No: 26 is the nucleotide sequence of the _VH region of antibody-024.

SEQ ID No：27是抗体-024 V_H区的氨基酸序列。SEQ ID No: 27 is the amino acid sequence of the _VH region of Antibody-024.

SEQ ID No：28是抗体-024 V_H CDR1的氨基酸序列，包括SEQ IDNo：27的31-35位氨基酸。SEQ ID No: 28 is the amino acid sequence of antibody-024 V _H CDR1, including amino acids 31-35 of SEQ ID No: 27.

SEQ ID No：29是抗体-024 V_H CDR2的氨基酸序列，包括SEQ IDNo：27的50-66位氨基酸。SEQ ID No: 29 is the amino acid sequence of Antibody-024 V _H CDR2, including amino acids 50-66 of SEQ ID No: 27.

SEQ ID No：30是抗体-024 V_H CDR3的氨基酸序列，包括SEQ IDNo：27的99-111位氨基酸。SEQ ID No: 30 is the amino acid sequence of antibody-024 V _H CDR3, including amino acids 99-111 of SEQ ID No: 27.

SEQ ID No：31是人CD38的序列。SEQ ID No: 31 is the sequence of human CD38.

SEQ ID No：32是突变型人CD38序列，其中237位的苏氨酸被丙氨酸替代。SEQ ID No: 32 is a mutant human CD38 sequence, in which threonine at position 237 is replaced by alanine.

SEQ ID No：33是突变型人CD38序列，其中272位的谷氨酸被精氨酸替代。SEQ ID No: 33 is a mutant human CD38 sequence, wherein glutamic acid at position 272 is replaced by arginine.

SEQ ID No：34是突变型人CD38序列，其中274位的丝氨酸被苯丙氨酸替代。SEQ ID No: 34 is a mutant human CD38 sequence, wherein the serine at position 274 is replaced by phenylalanine.

发明的详细描述Detailed description of the invention

本发明提供结合CD38的肽(“CD38BPs”)，它可用于治疗、诊断和预防各种由表达CD38的细胞参与的疾病，例如多发性骨髓瘤。The present invention provides CD38-binding peptides ("CD38BPs") that are useful in the treatment, diagnosis and prevention of various diseases involving CD38-expressing cells, such as multiple myeloma.

在一个技术方案中，本发明的CD38BP是抗体-003。-003是人单克隆IgG1抗体，具有包括SEQ ID No：2的序列的V_L区和包括SEQ ID No：7的序列的V_H区。In one technical solution, the CD38BP of the present invention is Antibody-003. -003 is a human monoclonal IgG1 antibody having a _VL region comprising the sequence of SEQ ID No:2 and a _VH region comprising the sequence of SEQ ID No:7.

在一个技术方案中，本发明的CD38BP是抗体-005。-005是人单克隆IgG1抗体，具有包括SEQ ID No：12的序列的V_L区和包括SEQ IDNo：17的序列的V_H区。In one technical solution, the CD38BP of the present invention is Antibody-005. -005 is a human monoclonal IgG1 antibody having a _VL region comprising the sequence of SEQ ID No: 12 and a _VH region comprising the sequence of SEQ ID No: 17.

在一个技术方案中，本发明的CD38BP是抗体-024。-024是人单克隆IgG1抗体，具有包括SEQ ID No：22的序列的V_L区和包括SEQ IDNo：27的序列的V_H区。In one technical solution, the CD38BP of the present invention is Antibody-024. -024 is a human monoclonal IgG1 antibody having a _VL region comprising the sequence of SEQ ID No:22 and a _VH region comprising the sequence of SEQ ID No:27.

抗体主要通过位于6个重链和轻链互补决定区(CDRs)的氨基酸残基与靶抗原相互作用。因此，在各个抗体间，CDRs中的氨基酸序列比CDRs之外的序列变异更多。由于CDR序列负责大部分抗体-抗原相互作用，因此可通过将来自特定天然存在的抗体CDR序列移植到来自具有不同特性的不同抗体的同框序列中而构建的表达载体来表达可模拟特定天然存在的抗体的性质的重组抗体(参见例如Riechmann，L.etal.，Nature 332，323-327(1998)、Jones，P.et al.，Nature 321，522-525(1986)and Queen，C.et al.，PNAS USA 86，10029-10033(1989))。Antibodies interact with target antigens primarily through amino acid residues located in the six heavy and light chain complementarity determining regions (CDRs). Thus, the amino acid sequences within the CDRs vary more among individual antibodies than sequences outside the CDRs. Since CDR sequences are responsible for most antibody-antigen interactions, expression vectors that can mimic specific naturally occurring antibodies can be expressed by grafting CDR sequences from a specific naturally occurring antibody into in-frame sequences from a different antibody with different properties. The recombinant antibody of the property of the antibody (see for example Riechmann, L. et al., Nature 332, 323-327 (1998), Jones, P. et al., Nature 321, 522-525 (1986) and Queen, C.et al., PNAS USA 86, 10029-10033 (1989)).

由于在本领域中已知抗体重链CDR3结构域在抗体与抗原的结合特异性/亲和性方面起非常重要的作用(Ditzel HJ，et al.，J Immunol.157(2)，739-49(1996)、Barbas SMet al.，J.Am.Chem.Soc.116，2161-2162(1994)，和Barbas SM et al.，Proc Natl AcadSci USA 92(7)，2529-33(1995))，因此本发明的CD3 8BPs可包括-003或-005或-024的重链CDR3s。本发明的CD38BPs也包括-003或-005或-024的轻链CDR3s。本发明的CD38BPs进一步分别包括-003或-005或-024的重链CDR2s。本发明的CD38BPs进一步分别包括-003或-005或-024的重链CDR1s。Since it is known in the art that the antibody heavy chain CDR3 domain plays a very important role in the binding specificity/affinity of the antibody to the antigen (Ditzel HJ, et al., J Immunol. 157(2), 739-49 (1996), Barbas SM et al., J.Am.Chem.Soc. 116, 2161-2162(1994), and Barbas SM et al., Proc Natl AcadSci USA 92(7), 2529-33(1995)), Thus CD3 8BPs of the invention may comprise heavy chain CDR3s of -003 or -005 or -024. The CD38BPs of the invention also include light chain CDR3s of -003 or -005 or -024. The CD38BPs of the present invention further comprise heavy chain CDR2s of -003 or -005 or -024, respectively. The CD38BPs of the present invention further comprise heavy chain CDR1s of -003 or -005 or -024, respectively.

本发明提供CD38BPs，它可与-003竞争结合CD38。The present invention provides CD38BPs that compete with -003 for CD38 binding.

本发明提供CD38BPs，它可与-005竞争结合CD38。The present invention provides CD38BPs that compete with -005 for CD38 binding.

本发明提供CD38BPs，它可与-024竞争结合CD38。The present invention provides CD38BPs that compete with -024 for CD38 binding.

在一个技术方案中，竞争作用通过如实施例部分描述的ELISA测定。In one embodiment, competition is determined by ELISA as described in the Examples section.

在一个技术方案中，竞争作用通过如实施例部分描述的FACS测定。In one embodiment, competition is determined by FACS as described in the Examples section.

本发明提供与CD38抗原决定簇结合的CD38BP，该抗原决定簇也与-003或-005或-024特异结合。The present invention provides CD38BP that binds to a CD38 epitope that also specifically binds to -003 or -005 or -024.

本发明提供与-003或-005或-024具有基本相同的结合人CD38的特异结合特性的CD38BP。The present invention provides a CD38BP having substantially the same specific binding properties to human CD38 as -003 or -005 or -024.

本发明提供含主要由SEQ ID No：3组成的V_L CDR1的CD38BP。The present invention provides a CD38BP comprising a _VL CDR1 consisting essentially of SEQ ID No:3.

本发明提供含主要由SEQ ID No：4组成的V_L CDR2的CD38BP。The present invention provides a CD38BP comprising a _VL CDR2 consisting essentially of SEQ ID No:4.

本发明提供含主要由SEQ ID No：5组成的V_L CDR3的CD38BP。The present invention provides a CD38BP comprising a _VL CDR3 consisting essentially of SEQ ID No:5.

本发明提供含主要由SEQ ID No：5组成的V_L CDR3和主要由SEQID No：3组成的V_LCDR1的CD38BP。The present invention provides a CD38BP comprising a _VL CDR3 consisting essentially of SEQ ID No:5 and a _VL CDR1 consisting essentially of SEQ ID No:3.

本发明提供含主要由SEQ ID No：5组成的V_L CDR3和主要由SEQID No：4组成的V_LCDR2的CD38BP。The present invention provides a CD38BP comprising a _VL CDR3 consisting essentially of SEQ ID No:5 and a _VL CDR2 consisting essentially of SEQ ID No:4.

本发明提供含主要由SEQ ID No：5组成的V_L CDR3和主要由SEQID No：4组成的V_LCDR2和主要由SEQ ID No：3组成的V_L CDR1的CD38BP。The present invention provides a CD38BP comprising _VL CDR3 consisting essentially of SEQ ID No:5, _VL CDR2 essentially consisting of SEQ ID No:4, and _VL CDR1 essentially consisting of SEQ ID No:3.

本发明提供含主要由SEQ ID No：8组成的V_H CDR1的CD38BP。The present invention provides a CD38BP comprising a _VH CDR1 consisting essentially of SEQ ID No:8.

本发明提供含主要由SEQ ID No：9组成的V_H CDR2的CD38BP。The present invention provides a CD38BP comprising a _VH CDR2 consisting essentially of SEQ ID No:9.

本发明提供含主要由SEQ ID No：10组成的V_H CDR3的CD38BP。The present invention provides a CD38BP comprising a _VH CDR3 consisting essentially of SEQ ID No:10.

本发明提供含主要由SEQ ID No：10组成的V_H CDR3和主要由SEQID No：8组成的V_HCDR1的CD38BP。The present invention provides a CD38BP comprising a _VH CDR3 consisting essentially of SEQ ID No: 10 and a _VH CDR1 consisting essentially of SEQ ID No:8.

本发明提供含主要由SEQ ID No：10组成的V_H CDR3和主要由SEQID No：9组成的V_HCDR2的CD38BP。The present invention provides a CD38BP comprising a _VH CDR3 consisting essentially of SEQ ID No: 10 and a _VH CDR2 consisting essentially of SEQ ID No:9.

本发明提供含主要由SEQ ID No：10组成的V_H CDR3和主要由SEQID No：9组成的V_HCDR2和主要由SEQ ID No：8组成的V_H CDR1的CD38BP。The present invention provides a CD38BP comprising a _VH CDR3 consisting essentially of SEQ ID No: 10, a _VH CDR2 consisting essentially of SEQ ID No: 9, and a _VH CDR1 consisting essentially of SEQ ID No: 8.

本发明提供含主要由SEQ ID No：13组成的V_L CDR1的CD38BP。The present invention provides a CD38BP comprising a _VL CDR1 consisting essentially of SEQ ID No:13.

本发明提供含主要由SEQ ID No：14组成的V_L CDR2的CD38BP。The present invention provides a CD38BP comprising a _VL CDR2 consisting essentially of SEQ ID No:14.

本发明提供含主要由SEQ ID No：15组成的V_L CDR3的CD38BP。The present invention provides a CD38BP comprising a _VL CDR3 consisting essentially of SEQ ID No:15.

本发明提供含主要由SEQ ID No：15组成的V_L CDR3和主要由SEQID No：13组成的V_LCDR1的CD38BP。The present invention provides a CD38BP comprising a V _L CDR3 consisting essentially of SEQ ID No: 15 and a V _L CDR1 consisting essentially of SEQ ID No: 13.

本发明提供含主要由SEQ ID No：15组成的V_L CDR3和主要由SEQID No：14组成的V_LCDR2的CD38BP。The present invention provides a CD38BP comprising a V _L CDR3 consisting essentially of SEQ ID No: 15 and a V _L CDR2 consisting essentially of SEQ ID No: 14.

本发明提供含主要由SEQ ID No：15组成的V_L CDR3和主要由SEQID No：14组成的V_LCDR2和主要由SEQ ID No：13组成的V_L CDR1的CD38BP。The present invention provides a CD38BP comprising a V _L CDR3 consisting essentially of SEQ ID No: 15, a V _L CDR2 consisting essentially of SEQ ID No: 14, and a V _L CDR1 consisting essentially of SEQ ID No: 13.

本发明提供含主要由SEQ ID No：18组成的V_H CDR1的CD38BP。The present invention provides a CD38BP comprising a _VH CDR1 consisting essentially of SEQ ID No:18.

本发明提供含主要由SEQ ID No：19组成的V_H CDR2的CD38BP。The present invention provides a CD38BP comprising a _VH CDR2 consisting essentially of SEQ ID No:19.

本发明提供含主要由SEQ ID No：20组成的V_H CDR3的CD38BP。The present invention provides a CD38BP comprising a _VH CDR3 consisting essentially of SEQ ID No:20.

本发明提供含主要由SEQ ID No：20组成的V_H CDR3和主要由SEQID No：18组成的V_HCDR1的CD38BP。The present invention provides a CD38BP comprising a _VH CDR3 consisting essentially of SEQ ID No:20 and a _VH CDR1 consisting essentially of SEQ ID No:18.

本发明提供含主要由SEQ ID No：20组成的V_H CDR3和主要由SEQID No：19组成的V_HCDR2的CD38BP。The present invention provides a CD38BP comprising a _VH CDR3 consisting essentially of SEQ ID No: 20 and a _VH CDR2 consisting essentially of SEQ ID No: 19.

本发明提供含主要由SEQ ID No：20组成的V_H CDR3和主要由SEQID No：19组成的V_HCDR2和主要由SEQ ID No：18组成的V_H CDR1的CD38BP。The present invention provides a CD38BP comprising a _VH CDR3 consisting essentially of SEQ ID No: 20, a _VH CDR2 consisting essentially of SEQ ID No: 19, and a _VH CDR1 consisting essentially of SEQ ID No: 18.

本发明提供含主要由SEQ ID No：23组成的V_L CDR1的CD38BP。The present invention provides a CD38BP comprising a _VL CDR1 consisting essentially of SEQ ID No:23.

本发明提供含主要由SEQ ID No：24组成的V_L CDR2的CD38BP。The present invention provides a CD38BP comprising a _VL CDR2 consisting essentially of SEQ ID No:24.

本发明提供含主要由SEQ ID No：25组成的V_L CDR3的CD3 8BP。The present invention provides a CD3 8BP comprising a _VL CDR3 consisting essentially of SEQ ID No:25.

本发明提供含主要由SEQ ID No：25组成的V_L CDR3和主要由SEQID No：23组成的V_LCDR1的CD38BP。The present invention provides a CD38BP comprising a V _L CDR3 consisting essentially of SEQ ID No: 25 and a V _L CDR1 consisting essentially of SEQ ID No: 23.

本发明提供含主要由SEQ ID No：25组成的V_L CDR3和主要由SEQID No：24组成的V_LCDR2的CD38BP。The present invention provides a CD38BP comprising a V _L CDR3 consisting essentially of SEQ ID No: 25 and a V _L CDR2 consisting essentially of SEQ ID No: 24.

本发明提供含主要由SEQ ID No：25组成的V_L CDR3和主要由SEQID No：24组成的V_LCDR2和主要由SEQ ID No：23组成的V_L CDR1的CD38BP。The present invention provides a CD38BP comprising a V _L CDR3 consisting essentially of SEQ ID No: 25, a V _L CDR2 consisting essentially of SEQ ID No: 24, and a V _L CDR1 consisting essentially of SEQ ID No: 23.

本发明提供含主要由SEQ ID No：28组成的V_H CDR1的CD38BP。The present invention provides a CD38BP comprising a _VH CDR1 consisting essentially of SEQ ID No:28.

本发明提供含主要由SEQ ID No：29组成的V_H CDR2的CD38BP。The present invention provides a CD38BP comprising a _VH CDR2 consisting essentially of SEQ ID No:29.

本发明提供含主要由SEQ ID No：30组成的V_H CDR3的CD38BP。The present invention provides a CD38BP comprising a _VH CDR3 consisting essentially of SEQ ID No:30.

本发明提供含主要由SEQ ID No：30组成的V_H CDR3和主要由SEQID No：28组成的V_HCDR1的CD38BP。The present invention provides a CD38BP comprising a _VH CDR3 consisting essentially of SEQ ID No:30 and a _VH CDR1 consisting essentially of SEQ ID No:28.

本发明提供含主要由SEQ ID No：30组成的V_H CDR3和主要由SEQID No：29组成的V_HCDR2的CD38BP。The present invention provides a CD38BP comprising a _VH CDR3 consisting essentially of SEQ ID No:30 and a _VH CDR2 consisting essentially of SEQ ID No:29.

本发明提供含主要由SEQ ID No：30组成的V_H CDR3和主要由SEQID No：29组成的V_HCDR2和主要由SEQ ID No：28组成的V_H CDR1的CD38BP。The present invention provides a CD38BP comprising a _VH CDR3 consisting essentially of SEQ ID No: 30, a _VH CDR2 consisting essentially of SEQ ID No: 29, and a _VH CDR1 consisting essentially of SEQ ID No: 28.

本发明提供CD38BP，包含The present invention provides CD38BP, comprising

(a)含有3个V_L CDRs的第一个V_L区，它们彼此间各自主要由SEQ ID No：3、SEQ IDNo：4和SEQ ID No：5组成；及(a) a first _VL region comprising 3 _VL CDRs, each of which consists essentially of SEQ ID No: 3, SEQ ID No: 4 and SEQ ID No: 5; and

(b)含有3个V_H CDRS的第一个V_H区，它们彼此间各自主要由SEQ ID No：8、SEQ IDNo：9和SEQ ID No：10组成。(b) The first _VH region containing 3 _VH CDRSs, which consist essentially of SEQ ID No: 8, SEQ ID No: 9 and SEQ ID No: 10 among themselves.

本发明提供CD38BP，包含The present invention provides CD38BP, comprising

(a)含有3个V_L CDRs的第一个V_L区，它们彼此间各自主要由SEQ ID No：13、SEQ IDNo：14和SEQ ID No：15组成；及(a) a first _VL region comprising 3 _VL CDRs, each of which consists essentially of SEQ ID No: 13, SEQ ID No: 14 and SEQ ID No: 15; and

(b)含有3个V_H CDRs的第一个V_H区，它们彼此间各自主要由SEQ ID No：18、SEQ IDNo：19和SEQ ID No：20组成。(b) The first _VH region containing 3 _VH CDRs, which consist essentially of SEQ ID No: 18, SEQ ID No: 19 and SEQ ID No: 20 each other.

本发明提供CD38BP，包含The present invention provides CD38BP, comprising

(a)含有3个V_L CDRs的第一个V_L区，它们彼此间各自主要由SEQ ID No：23、SEQ IDNo：24和SEQ ID No：25组成；及(a) a first _VL region comprising 3 _VL CDRs, each of which consists essentially of SEQ ID No: 23, SEQ ID No: 24 and SEQ ID No: 25; and

(b)含有3个V_H CDRS的第一个V_H区，它们彼此间各自主要由SEQ ID No：28、SEQ IDNo：29和SEQ ID No：30组成。(b) The first _VH region containing 3 _VH CDRSs, which consist essentially of SEQ ID No: 28, SEQ ID No: 29 and SEQ ID No: 30 each other.

在一个技术方案中，V_L区和V_H区位于肽的同一条链上。In one technical solution, the _VL and _VH regions are located on the same chain of the peptide.

在另一个技术方案中，V_L区和V_H区由柔性的连接物分开。In another embodiment, the _VL and _VH domains are separated by a flexible linker.

在一个技术方案中，V_L区和V_H区位于肽的单独的链上。In one embodiment, the _VL and _VH regions are located on separate chains of the peptide.

在另一个技术方案中，V_L区和V_H区位于作为免疫球蛋白折叠蛋白的肽的不同链上。In another embodiment, the _VL and _VH regions are located on different chains of a peptide that is a folded immunoglobulin protein.

在一个技术方案中，第一个V_L区和第一个V_H区是定向的，从而使V_L区的3个CDRs和V_H区的3个CDRs协同作用来选择性地和/或特异性地与人CD38上的抗原决定簇结合。In one technical solution, the first _VL region and the first _VH region are oriented so that the 3 CDRs of the _VL region and the 3 CDRs of the _VH region cooperate to selectively and/or specifically Binds positively to an epitope on human CD38.

在另一个技术方案中，肽包括第二个与第一个V_L区相同的V_L区和第二个与第一个V_H区相同的V_H区，其中第二个V_L区和第二个V_H区一起用于选择性地和/或特异性地结合人CD38的抗原决定簇。In another technical solution, the peptide comprises a second _VL region identical to the first _VL region and a second _VH region identical to the first _VH region, wherein the second _VL region and the second Together, the two _VH regions serve to selectively and/or specifically bind epitopes of human CD38.

本发明提供含有作为-003或-005或-024的V_L区的功能变异体的V_L 区的CD38BP。The present invention provides CD38BPs containing a _VL region that is a functional variant of the _VL region of -003 or -005 or -024.

在一个技术方案中，CD38BP的V_L区主要由与分别如SEQ ID No：2或SEQ ID No：12或SEQ ID No：22中所述序列的氨基酸同一性至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％的序列组成。在一个技术方案中，CD38BP分别具有至少约50％、至少约60％、至少约70％、至少约80％、至少约90％或至少约95％的-003或-005或-024的抗原决定簇结合特性。In one technical solution, the _VL region of CD38BP consists essentially of at least about 50%, at least about 60% amino acid identity to the sequence set forth in SEQ ID No: 2 or SEQ ID No: 12 or SEQ ID No: 22, respectively. , at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% sequence composition. In one technical solution, CD38BP has at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 95% of the antigenic determination of -003 or -005 or -024, respectively Cluster binding properties.

本发明提供含有作为-003或-005或-024的V_H区的功能变异体的V_H 区的CD38BP。The present invention provides CD38BPs containing a _VH region that is a functional variant of the _VH region of -003 or -005 or -024.

在一个技术方案中，CD3 8BP的V_H区主要由与分别如SEQ ID No：7或SEQ ID No：17或SEQ ID No：27中所述序列的氨基酸同一性至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％的序列组成。在一个技术方案中，CD38BP分别具有至少约50％、至少约60％、至少约70％、至少约80％、至少约90％或至少约95％的-003或-005或-024的抗原决定簇结合特性。In one technical solution, the _VH region of CD3 8BP consists essentially of at least about 50%, at least about 60% amino acid identity to the sequence set forth in SEQ ID No: 7 or SEQ ID No: 17 or SEQ ID No: 27, respectively. %, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% sequence composition. In one technical solution, CD38BP has at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 95% of the antigenic determination of -003 or -005 or -024, respectively Cluster binding properties.

本发明提供含至少一个作为-003或-005或-024的CDR的功能变异体的CDR的CD38BP。The present invention provides a CD38BP comprising at least one CDR that is a functional variant of the CDR of -003 or -005 or -024.

在一个技术方案中，至少一个肽的CDRs主要由与分别如SEQ IDNo：3、SEQ ID No：4、SEQ ID No：5、SEQ ID No：8、SEQ ID No：9或SEQ ID No：10所述，或如SEQ ID No：13、SEQID No：14、SEQ IDNo：15、SEQ ID No：18、SEQ ID No：19，或如SEQ ID No：20所述或如SEQ IDNo：23、SEQ ID No：24、SEQ ID No：25、SEQ ID No：28、SEQ ID No：29或SEQ ID No：30中所述序列的氨基酸序列同一性至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％的序列组成。在一个技术方案中，CD38BP分别具有至少约50％、至少约60％、至少约70％、至少约80％、至少约90％或至少约95％的-003或-005或-024的抗原决定簇结合特性。In a technical solution, the CDRs of at least one peptide are mainly composed of peptides such as SEQ ID No: 3, SEQ ID No: 4, SEQ ID No: 5, SEQ ID No: 8, SEQ ID No: 9 or SEQ ID No: 10 Described, or as SEQ ID No: 13, SEQ ID No: 14, SEQ ID No: 15, SEQ ID No: 18, SEQ ID No: 19, or as described in SEQ ID No: 20 or as SEQ ID No: 23, SEQ ID No: 23, SEQ ID No: 19, or as described in SEQ ID No: 20 The amino acid sequence identity of the sequence set forth in ID No: 24, SEQ ID No: 25, SEQ ID No: 28, SEQ ID No: 29 or SEQ ID No: 30 is at least about 50%, at least about 60%, at least about 70%. %, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% sequence composition. In one technical solution, CD38BP has at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 95% of the antigenic determination of -003 or -005 or -024, respectively Cluster binding properties.

在一个技术方案中，CD38BP分别具有至少约50％、至少约60％、至少约70％、至少约80％、至少约90％或至少约95％的-003或-005或-024的亲和性、强度或特异性。In one technical solution, the CD38BP has at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 95% of the affinity of -003 or -005 or -024, respectively sex, intensity or specificity.

在一个技术方案中，CD38BP与-003或-005或-024竞争结合CD38。In one technical solution, CD38BP competes with -003 or -005 or -024 for binding to CD38.

在另一个技术方案中，竞争作用通过如实施例部分所述的ELISA进 行测定。在另一个技术方案中，竞争作用通过如实施例部分所述的FACS进行测定。In another embodiment, competition is determined by ELISA as described in the Examples section. In another embodiment, competition is determined by FACS as described in the Examples section.

在一个技术方案中，CD38BP与CD38抗原决定簇特异结合，其抗原决定簇也与-003或-005或-024特异结合。In a technical scheme, CD38BP specifically binds to the CD38 antigenic determinant, and its antigenic determinant also specifically binds to -003 or -005 or -024.

在一个技术方案中，CD38BP与人CD38结合的亲和性大于-003或-005或-024。In one technical solution, the binding affinity of CD38BP to human CD38 is greater than -003 or -005 or -024.

在一个技术方案中，CD38BP实质具有与-003或-005或-024相同的特异结合CD38的特性。In a technical solution, CD38BP substantially has the same characteristic of specifically binding to CD38 as -003 or -005 or -024.

在一个技术方案中，CD38BP实质没有其它CD38结合肽。In one embodiment, the CD38BP is substantially free of other CD38 binding peptides.

在一个技术方案中，本发明的CD38BP是抗体。在另一个技术方案中，CD38BP是人抗体。在另一个技术方案中，CD38BP是人源化抗体。在另一个技术方案中，CD38BP是嵌合抗体。In one technical solution, the CD38BP of the present invention is an antibody. In another embodiment, CD38BP is a human antibody. In another technical solution, CD38BP is a humanized antibody. In another technical solution, CD38BP is a chimeric antibody.

在一个技术方案中，本发明的抗体是单克隆抗体。In one embodiment, the antibody of the invention is a monoclonal antibody.

在一个技术方案中，本发明的抗体是IgG1、IgG2、IgG3、IgG4、IgD、IgA、IgE、或IgM抗体。在另一个技术方案中，抗体是IgG1抗体。在另一个技术方案中，抗体是IgG1κ抗体。在另一个技术方案中，抗体是IgM抗体。在另一个技术方案中，抗体是IgM1κ抗体。In one technical solution, the antibody of the present invention is an IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE, or IgM antibody. In another embodiment, the antibody is an IgG1 antibody. In another embodiment, the antibody is an IgG1κ antibody. In another embodiment, the antibody is an IgM antibody. In another embodiment, the antibody is an IgM1κ antibody.

在一个技术方案中，本发明的抗体是抗体片段或单链抗体。In one technical solution, the antibody of the present invention is an antibody fragment or a single chain antibody.

在一个技术方案中，CD38BP在真核细胞中是糖基化的。In one embodiment, CD38BP is glycosylated in eukaryotic cells.

在一个技术方案中，CD38BP进一步含有用于连接放射性同位素的螯合剂连接物。In one technical solution, CD38BP further contains a chelator linker for attaching a radioisotope.

在一个技术方案中，CD38BP实质是分离的形式。In one embodiment, CD38BP is substantially in isolated form.

本发明提供分离的编码本发明的CD38BP的核酸。The invention provides an isolated nucleic acid encoding a CD38BP of the invention.

本发明提供含编码本发明的CD38BP的核酸序列的表达载体。The present invention provides an expression vector containing the nucleic acid sequence encoding CD38BP of the present invention.

在一个技术方案中，表达载体含序列号为SEQ ID No：1的V_L核苷酸序列，序列号为SEQ ID No：6的V_H核苷酸序列，或序列号为SEQ IDNo：1的V_L核苷酸序列和序列号为SEQ IDNo：6的V_H核苷酸序列。In one technical solution, the expression vector contains the V _L nucleotide sequence whose sequence number is SEQ ID No: 1, the V _H nucleotide sequence whose sequence number is SEQ ID No: 6, or the sequence number that is SEQ ID No: 1 The V _L nucleotide sequence and the V _H nucleotide sequence whose sequence number is SEQ ID No: 6.

在一个技术方案中，表达载体含序列号为SEQ ID No：11的V_L核苷酸序列，序列号为SEQ ID No：16的V_H核苷酸序列，或序列号为SEQID No：11的V_L核苷酸序列和序列号为SEQID No：16的V_H核苷酸序列。In one technical solution, the expression vector contains the _VL nucleotide sequence whose sequence number is SEQ ID No: 11, the _VH nucleotide sequence whose sequence number is SEQ ID No: 16, or the sequence number that is SEQID No: 11 The V _L nucleotide sequence and the V _H nucleotide sequence whose sequence number is SEQID No: 16.

在一个技术方案中，表达载体含序列号为SEQ ID No：21的V_L核 苷酸序列，序列号为SEQ ID No：26的V_H核苷酸序列，或序列号为SEQID No：21的V_L核苷酸序列和序列号为SEQID No：26的V_H核苷酸序列。In one technical solution, the expression vector contains the _VL nucleotide sequence whose sequence number is SEQ ID No: 21, the _VH nucleotide sequence whose sequence number is SEQ ID No: 26, or the sequence number that is SEQID No: 21 The V _L nucleotide sequence and the V _H nucleotide sequence whose sequence number is SEQID No: 26.

在另一个技术方案中，表达载体进一步含编码人抗体的轻链、重链或轻链和重链恒定区的核苷酸序列。In another technical solution, the expression vector further comprises a nucleotide sequence encoding the constant regions of the light chain, the heavy chain, or the light chain and the heavy chain of a human antibody.

在另一个技术方案中，编码人抗体的轻链、重链或轻链和重链恒定区的核苷酸序列编码IgG1抗体。In another technical solution, the nucleotide sequence encoding the light chain, the heavy chain, or the light and heavy chain constant regions of a human antibody encodes an IgG1 antibody.

本发明提供可产生由人轻链和人重链核酸编码的人单克隆抗CD38抗体的杂交瘤，这些序列包括如SEQ ID No：1中的可变轻链区的核苷酸序列或其保守序列改变，及如SEQID No：6中的可变重链区的核苷酸序列或其保守序列改变。在一个技术方案中，人轻链核酸含有如SEQID No：1所示的核苷酸序列，人重链核酸含有如SEQ ID No：6所示的核苷酸序列。The present invention provides hybridomas capable of producing human monoclonal anti-CD38 antibodies encoded by human light chain and human heavy chain nucleic acids, which sequences include the nucleotide sequence of the variable light chain region as in SEQ ID No: 1 or its conserved Sequence changes, and changes in the nucleotide sequence of the variable heavy chain region as in SEQ ID No: 6 or its conservative sequence. In one technical solution, the human light chain nucleic acid contains the nucleotide sequence shown in SEQ ID No: 1, and the human heavy chain nucleic acid contains the nucleotide sequence shown in SEQ ID No: 6.

本发明提供可产生具有人轻链和重链可变区的人单克隆抗CD38抗体的杂交瘤，它包括如SEQ ID No：2或其保守序列改变中的人轻链可变氨基酸序列，及如SEQ ID No：7或其保守序列改变中的人重链可变氨基酸序列。在一个技术方案中，人轻链可变区含有如SEQID No：2中的氨基酸序列，人重链可变区含有如SEQ ID No：7中的氨基酸序列。The present invention provides a hybridoma capable of producing a human monoclonal anti-CD38 antibody having human light chain and heavy chain variable regions, comprising the human light chain variable amino acid sequence as in SEQ ID No: 2 or a conservative sequence alteration thereof, and Human heavy chain variable amino acid sequence as in SEQ ID No: 7 or conservative sequence variations thereof. In one technical solution, the human light chain variable region contains the amino acid sequence as in SEQ ID No: 2, and the human heavy chain variable region contains the amino acid sequence as in SEQ ID No: 7.

本发明提供可产生由如SEQ ID No：1中的人轻链可变区的核酸或其保守序列改变及如SEQ ID No：6中的人重链核酸或其保守序列改变编码的人单克隆抗CD38抗体的转染瘤。在一个技术方案中，人单克隆抗CD38抗体由如SEQ ID No：1中的人轻链可变区核酸和如SEQ IDNo：6中的人重链核酸编码。The present invention provides human monoclonals that can be produced by the nucleic acid of the human light chain variable region as in SEQ ID No: 1 or its conserved sequence change and the human heavy chain nucleic acid as in SEQ ID No: 6 or its conserved sequence change Transfectomas with anti-CD38 antibodies. In one technical solution, the human monoclonal anti-CD38 antibody is encoded by the human light chain variable region nucleic acid as in SEQ ID No:1 and the human heavy chain nucleic acid as in SEQ ID No:6.

本发明提供可产生具有人轻链和重链可变区的人单克隆抗CD38抗体的转染瘤，这些可变区包括如SEQ ID No：2中的人轻链可变区的氨基酸序列或其保守序列改变，及如SEQID No：7中的人重链可变区的氨基酸序列或其保守序列改变。在一个技术方案中，人轻链含有如SEQID No：2中的人轻链可变区氨基酸序列，人重链含有如SEQ ID No：7中的人重链可变区氨基酸序列。The present invention provides a transfectoma that produces a human monoclonal anti-CD38 antibody having human light and heavy chain variable regions comprising the amino acid sequence of the human light chain variable region in SEQ ID No: 2 or Its conservative sequence changes, and the amino acid sequence of the human heavy chain variable region in SEQID No: 7 or its conservative sequence changes. In one technical solution, the human light chain contains the amino acid sequence of the human light chain variable region as in SEQ ID No: 2, and the human heavy chain contains the amino acid sequence of the human heavy chain variable region as in SEQ ID No: 7.

本发明提供可产生由人轻链和人重链核酸编码的人单克隆抗CD38抗体的杂交瘤，这些序列包括如SEQ ID No：11中的可变轻链区的核苷酸序列或其保守序列改变，及如SEQID No：16中的可变重链区的核苷 酸序列或其保守序列改变。在一个技术方案中，人轻链核酸含有如SEQID No：11所示的核苷酸序列，人重链核酸含有如SEQ ID No：16所示的核苷酸序列。The present invention provides hybridomas capable of producing human monoclonal anti-CD38 antibodies encoded by human light chain and human heavy chain nucleic acids, which sequences include the nucleotide sequence of the variable light chain region as in SEQ ID No: 11 or its conserved Sequence changes, and changes in the nucleotide sequence of the variable heavy chain region as in SEQ ID No: 16 or its conservative sequence. In one technical solution, the human light chain nucleic acid contains the nucleotide sequence shown in SEQ ID No: 11, and the human heavy chain nucleic acid contains the nucleotide sequence shown in SEQ ID No: 16.

本发明提供可产生具有人重链和轻链可变区的人单克隆抗CD38抗体的杂交瘤，这些可变区包括如SEQ ID No：12中的人轻链可变区的氨基酸序列或其保守序列改变，及如SEQ ID No：17中的人重链可变区的氨基酸序列或其保守序列改变。在一个技术方案中，人轻链含有如SEQID No：12中的人轻链可变区氨基酸序列，人重链含有如SEQ ID No：17中的人重链可变区氨基酸序列。The present invention provides a hybridoma capable of producing a human monoclonal anti-CD38 antibody having human heavy and light chain variable regions comprising the amino acid sequence of the human light chain variable region as in SEQ ID No: 12 or its Conservative sequence changes, and the amino acid sequence of the human heavy chain variable region as in SEQ ID No: 17 or its conservative sequence changes. In one technical solution, the human light chain contains the amino acid sequence of the human light chain variable region as in SEQ ID No: 12, and the human heavy chain contains the amino acid sequence of the human heavy chain variable region as in SEQ ID No: 17.

本发明提供可产生由如SEQ ID No：11中的人轻链可变区的核酸或其保守序列改变及如SEQ ID No：16中的人重链核酸或其保守序列改变编码的人单克隆抗CD38抗体的转染瘤。在一个技术方案中，人单克隆抗CD38抗体由如SEQ ID No：11中的人轻链可变区核酸和如SEQ IDNo：16中的人重链核酸编码。The present invention provides a human monoclonal that can be produced by the nucleic acid of the human light chain variable region as in SEQ ID No: 11 or its conserved sequence change and the human heavy chain nucleic acid as in SEQ ID No: 16 or its conserved sequence change Transfectomas with anti-CD38 antibodies. In one technical solution, the human monoclonal anti-CD38 antibody is encoded by the human light chain variable region nucleic acid as in SEQ ID No: 11 and the human heavy chain nucleic acid as in SEQ ID No: 16.

本发明提供可产生具有人轻链和重链可变区的人单克隆抗CD38抗体的转染瘤，这些可变区包括如SEQ ID No：12中的人轻链可变区的氨基酸序列或其保守序列改变，及如SEQ ID No：17中的人重链可变区的氨基酸序列或其保守序列改变。在一个技术方案中，人轻链含有如SEQID No：12中的人轻链可变区氨基酸序列，人重链含有如SEQ ID No：17中的人重链可变区氨基酸序列。The present invention provides a transfectoma that produces a human monoclonal anti-CD38 antibody having human light and heavy chain variable regions comprising the amino acid sequence of the human light chain variable region in SEQ ID No: 12 or Its conservative sequence changes, and the amino acid sequence of the human heavy chain variable region as in SEQ ID No: 17 or its conservative sequence changes. In one technical solution, the human light chain contains the amino acid sequence of the human light chain variable region as in SEQ ID No: 12, and the human heavy chain contains the amino acid sequence of the human heavy chain variable region as in SEQ ID No: 17.

本发明提供可产生由人轻链和人重链核酸编码的人单克隆抗CD38抗体的杂交瘤，这些序列包括如SEQ ID No：21中的可变轻链区的核苷酸序列或其保守序列改变，及如SEQID No：26中的可变重链区的核苷酸序列或其保守序列改变。在一个技术方案中，人轻链核酸含有如SEQID No：21所示的核苷酸序列，人重链核酸含有如SEQ ID No：26所示的核苷酸序列。The present invention provides hybridomas that can produce human monoclonal anti-CD38 antibodies encoded by human light chain and human heavy chain nucleic acids, these sequences include the nucleotide sequence of the variable light chain region as in SEQ ID No: 21 or its conserved Sequence changes, and changes in the nucleotide sequence of the variable heavy chain region as in SEQ ID No: 26 or its conservative sequence. In one technical solution, the human light chain nucleic acid contains the nucleotide sequence shown in SEQ ID No: 21, and the human heavy chain nucleic acid contains the nucleotide sequence shown in SEQ ID No: 26.

本发明提供可产生具有人重链和轻链可变区的人单克隆抗CD38抗体的转染瘤，这些可变区包括如SEQ ID No：22中的人轻链可变区的氨基酸序列或其保守序列改变，及如SEQ ID No：27中的人重链可变区的氨基酸序列或其保守序列改变。在一个技术方案中，人轻链可变区含有如SEQ ID No：22中的人轻链可变区氨基酸序列，人重链可变区含有如SEQID No：27中的人重链可变区氨基酸序列。The present invention provides a transfectoma that produces a human monoclonal anti-CD38 antibody having human heavy and light chain variable regions comprising the amino acid sequence of the human light chain variable region as in SEQ ID No: 22 or Its conservative sequence changes, and the amino acid sequence of the human heavy chain variable region in SEQ ID No: 27 or its conservative sequence changes. In one technical solution, the human light chain variable region contains the amino acid sequence of the human light chain variable region as in SEQ ID No: 22, and the human heavy chain variable region contains the human heavy chain variable region as in SEQ ID No: 27 amino acid sequence.

本发明提供可产生由如SEQ ID No：21中的人轻链可变区的核酸或其保守序列改变及如SEQ ID No：26中的人重链核酸或其保守序列改变编码的人单克隆抗CD38抗体的转染瘤。在一个技术方案中，人单克隆抗CD3 8抗体由如SEQ ID No：21中的人轻链可变区核酸和如SEQ IDNo：26中的人重链核酸编码。The present invention provides human monoclonals that can be produced by the nucleic acid of the human light chain variable region as in SEQ ID No: 21 or its conserved sequence change and the human heavy chain nucleic acid as in SEQ ID No: 26 or its conserved sequence change Transfectomas with anti-CD38 antibodies. In one technical solution, the human monoclonal anti-CD38 antibody is encoded by the human light chain variable region nucleic acid as in SEQ ID No:21 and the human heavy chain nucleic acid as in SEQ ID No:26.

本发明提供可产生具有人轻链和重链可变区的人单克隆抗CD38抗体的转染瘤，这些可变区包括如SEQ ID No：22中的人轻链可变区的氨基酸序列或其保守序列改变，及如SEQ ID No：27中的人重链可变区的氨基酸序列或其保守序列改变。在一个技术方案中，人轻链含有如SEQID No：22中的人轻链可变区氨基酸序列，人重链含有如SEQ ID No：27中的人重链可变区氨基酸序列。The present invention provides a transfectoma that produces a human monoclonal anti-CD38 antibody having human light and heavy chain variable regions comprising the amino acid sequence of the human light chain variable region in SEQ ID No: 22 or Its conservative sequence changes, and the amino acid sequence of the human heavy chain variable region in SEQ ID No: 27 or its conservative sequence changes. In one technical solution, the human light chain contains the amino acid sequence of the human light chain variable region as in SEQ ID No: 22, and the human heavy chain contains the amino acid sequence of the human heavy chain variable region as in SEQ ID No: 27.

本发明提供可产生本发明的CD38BP的真核或原核宿主细胞。The invention provides eukaryotic or prokaryotic host cells capable of producing the CD38BP of the invention.

本发明提供含有本发明的表达载体的真核或原核宿主细胞。The invention provides eukaryotic or prokaryotic host cells containing the expression vectors of the invention.

本发明提供含有编码人重链和人轻链的核酸的转基因动物或植物，其中动物或植物产生可检测量的本发明的CD38BP。The invention provides a transgenic animal or plant comprising nucleic acid encoding a human heavy chain and a human light chain, wherein the animal or plant produces a detectable amount of a CD38BP of the invention.

本发明提供与腺病毒药剂、放射性同位素或药物连接的本发明的CD38BP的免疫连接物。在一个技术方案中，肽是与腺病毒药剂、放射性同位素或药物连接的单价的IgM抗体。The invention provides an immunoconjugate of the CD38BP of the invention linked to an adenoviral agent, radioisotope or drug. In one embodiment, the peptide is a monovalent IgM antibody linked to an adenoviral agent, radioisotope or drug.

本发明提供含有本发明的CD38BP，并对人效应细胞具有结合特异性的双特异或多重特异的分子。在一个技术方案中，对人效应细胞的结合特异性是指对CD3、CD4、CD138、IL-15R、膜结合或受体结合的TNF-α、人Fc受体或膜结合或受体结合的IL-15的结合特异性。The present invention provides bispecific or multispecific molecules containing CD38BP of the present invention and having binding specificity to human effector cells. In one technical solution, binding specificity to human effector cells refers to CD3, CD4, CD138, IL-15R, membrane bound or receptor bound TNF-α, human Fc receptor or membrane bound or receptor bound Binding specificity of IL-15.

本发明提供与本发明的CD38BP结合的抗独特型抗体。本发明提供本发明的抗独特型抗体在检测样品中抗CD38的人单克隆抗体水平方面的用途。The invention provides anti-idiotypic antibodies that bind to a CD38BP of the invention. The present invention provides the use of the anti-idiotypic antibody of the present invention in detecting the level of human monoclonal antibody against CD38 in a sample.

以下是本发明选择的技术方案的列表。The following is a list of technical solutions selected by the present invention.

技术方案1：与人CD38结合的抗体，由可变区分别含有如SEQ IDNo：1和SEQ ID No：6所示的核苷酸序列、或是其保守序列改变的人轻链和人重链核酸编码。Technical solution 1: An antibody that binds to human CD38, the variable regions contain the nucleotide sequences shown in SEQ ID No: 1 and SEQ ID No: 6, or human light chains and human heavy chains with conservative sequence changes nucleic acid code.

技术方案2：与人CD38结合的抗体，由可变区分别含有如SEQ IDNo：1和SEQ ID No：6所示的核苷酸序列的人轻链和人重链核酸编码。Technical solution 2: The antibody binding to human CD38 is encoded by human light chain and human heavy chain nucleic acids whose variable regions contain the nucleotide sequences shown in SEQ ID No: 1 and SEQ ID No: 6, respectively.

技术方案3：与人CD38结合的抗体，由可变区分别含有如SEQ ID No：11和SEQ IDNo：16所示的核苷酸序列、或是其保守序列改变的人轻链和人重链核酸编码。Technical scheme 3: The antibody binding to human CD38, the variable regions respectively contain the nucleotide sequences shown in SEQ ID No: 11 and SEQ ID No: 16, or human light chains and human heavy chains whose conservative sequences are changed nucleic acid code.

技术方案4：与人CD38结合的抗体，由可变区分别含有如SEQ IDNo：11和SEQ IDNo：26所示的核苷酸序列的人轻链和人重链核酸编码。Technical solution 4: The antibody binding to human CD38 is encoded by human light chain and human heavy chain nucleic acids whose variable regions contain the nucleotide sequences shown in SEQ ID No: 11 and SEQ ID No: 26, respectively.

技术方案5：与人CD38结合的抗体，由可变区分别含有如SEQ IDNo：21和SEQ IDNo：26所示的核苷酸序列、或是其保守序列改变的人轻链和人重链核酸编码。Technical scheme 5: The antibody binding to human CD38, the variable regions respectively contain the nucleotide sequences shown in SEQ ID No: 21 and SEQ ID No: 26, or the human light chain and human heavy chain nucleic acids whose conservative sequences are changed coding.

技术方案6：与人CD38结合的抗体，由可变区分别含有如SEQ IDNo：21和SEQ IDNo：26所示的核苷酸序列的人轻链和人重链核酸编码。Technical solution 6: The antibody binding to human CD38 is encoded by human light chain and human heavy chain nucleic acids whose variable regions contain the nucleotide sequences shown in SEQ ID No: 21 and SEQ ID No: 26, respectively.

技术方案7：与如技术方案2所述的抗体竞争结合CD38的肽。Technical scheme 7: A peptide that competes with the antibody described in technical scheme 2 for binding to CD38.

技术方案8：如技术方案7所述的肽，其中竞争作用是由说明书的实施例8或9中所述的ELISA测定的。Technical solution 8: The peptide according to technical solution 7, wherein the competition is determined by ELISA described in Example 8 or 9 of the specification.

技术方案9：如技术方案7所述的肽，其中竞争作用是由说明书的实施例7中所述的交叉抑制方法测定的。Technical Solution 9: The peptide according to Technical Solution 7, wherein the competition is determined by the cross-inhibition method described in Example 7 of the specification.

技术方案10：与CD38抗原决定簇特异结合的肽，该抗原决定簇也可被如技术方案2所述的抗体特异结合。Technical scheme 10: a peptide that specifically binds to a CD38 epitope, and the antigenic determinant can also be specifically bound by the antibody described in technical scheme 2.

技术方案11：与技术方案2所述的抗体实质具有相同的特异结合人CD38的结合特性的肽。Technical solution 11: a peptide having substantially the same binding property of specifically binding to human CD38 as the antibody described in technical solution 2.

技术方案12：含有主要由SEQ ID No：3组成的V_L CDR1的肽。Technical solution 12: A peptide containing V _L CDR1 mainly composed of SEQ ID No:3.

技术方案13：含有主要由SEQ ID No：4组成的V_L CDR2的肽。Technical solution 13: A peptide containing V _L CDR2 mainly composed of SEQ ID No:4.

技术方案14：含有主要由SEQ ID No：5组成的V_L CDR3的肽。Technical solution 14: A peptide containing V _L CDR3 mainly composed of SEQ ID No:5.

技术方案15：如技术方案14所述的肽，该肽也含有主要由SEQ IDNo：3组成的V_LCDR1。Technical solution 15: The peptide according to technical solution 14, which also contains V _L CDR1 mainly composed of SEQ ID No:3.

技术方案16：如技术方案14所述的肽，该肽也含有主要由SEQ IDNo：4组成的V_LCDR2。Technical solution 16: The peptide according to technical solution 14, which also contains V _L CDR2 mainly composed of SEQ ID No:4.

技术方案17：如技术方案16所述的肽，该肽也含有主要由SEQ IDNo：3组成的V_LCDR1。Technical scheme 17: The peptide according to technical scheme 16, which also contains V _L CDR1 mainly consisting of SEQ ID No:3.

技术方案18：含有主要由SEQ ID No：8组成的V_H CDR1的肽。Technical solution 18: A peptide containing _VH CDR1 mainly composed of SEQ ID No:8.

技术方案19：含有主要由SEQ ID No：9组成的V_H CDR2的肽。Technical solution 19: A peptide containing _VH CDR2 mainly composed of SEQ ID No:9.

技术方案20：含有主要由SEQ ID No：10组成的V_H CDR3的肽。Technical solution 20: A peptide containing _VH CDR3 mainly composed of SEQ ID No:10.

技术方案21：如技术方案20所述的肽，该肽也含有主要由SEQ IDNo：8组成的V_HCDR1。Technical solution 21: The peptide according to technical solution 20, which also contains _VH CDR1 mainly composed of SEQ ID No:8.

技术方案22：如技术方案20所述的肽，该肽也含有主要由SEQ IDNo：9组成的V_HCDR2。Technical solution 22: The peptide according to technical solution 20, which also contains _VH CDR2 mainly composed of SEQ ID No:9.

技术方案23：如技术方案22所述的肽，该肽也含有主要由SEQ IDNo：8组成的V_HCDR1。Technical solution 23: The peptide according to technical solution 22, which also contains _VH CDR1 mainly composed of SEQ ID No:8.

技术方案24：肽，含有Technical scheme 24: peptide, containing

(a)含有3个V_L CDRs的第一个V_L区，它们彼此间各自主要由SEQ ID No：3、SEQ IDNo：4和SEQ ID No：5组成；及(a) a first _VL region comprising 3 _VL CDRs, each of which consists essentially of SEQ ID No: 3, SEQ ID No: 4 and SEQ ID No: 5; and

(b)含有3个V_H CDRs的第一个V_H区，它们彼此间各自主要由SEQ ID No：8、SEQ IDNo：9和SEQ ID No：10组成。(b) A first _VH region containing 3 _VH CDRs, which consist essentially of SEQ ID No: 8, SEQ ID No: 9 and SEQ ID No: 10 with respect to each other.

技术方案25：如技术方案24所述的肽，其中V_L区和V_H区位于肽的同一条链上。Technical solution 25: The peptide according to technical solution 24, wherein the _VL region and the _VH region are located on the same chain of the peptide.

技术方案26：如技术方案25所述的肽，其中V_L区和V_H区由柔性的连接物分隔开。Technical solution 26: The peptide according to technical solution 25, wherein the _VL region and the _VH region are separated by a flexible linker.

技术方案27：如技术方案24所述的肽，其中V_L区和V_H区位于肽的不同的链上。Technical solution 27: The peptide according to technical solution 24, wherein the _VL region and the _VH region are located on different chains of the peptide.

技术方案28：如技术方案27所述的肽，其中V_L区和V_H区位于作为免疫球蛋白折叠蛋白的肽的不同链上。Technical solution 28: The peptide according to technical solution 27, wherein the _VL region and the _VH region are located on different chains of the peptide which is a folded immunoglobulin protein.

技术方案29：如技术方案24到28任何一项中所述的肽，其中第一个V_L区和第一个V_H区是定向的，从而使V_L区的3个CDRs和V_H区的3个CDRs协同作用来选择性地和/或特异性地与人CD38上的抗原决定簇结合。Technical scheme 29: The peptide as described in any one of technical scheme 24 to 28, wherein the first _VL district and the first _VH district are oriented, thereby make 3 CDRs of _VL district and _VH district The three CDRs of the CD38 work together to selectively and/or specifically bind to epitopes on human CD38.

技术方案30：如技术方案29所述的肽，其中肽含有第二个与第一个V_L区相同的V_L区和第二个与第一个V_H区相同的V_H区，其中第二个V_L区和第二个V_H区协同作用来选择性地和/或特异性地与人CD38上的抗原决定簇结合。Technical scheme 30: The peptide of technical scheme 29, wherein the peptide contains a second _VL region identical to the first _VL region and a second _VH region identical to the first _VH region, wherein the second The two _VL domains and the second _VH domain cooperate to selectively and/or specifically bind to antigenic determinants on human CD38.

技术方案31：含有作为技术方案2的抗体V_L区功能变异体的V_L 区的肽。Technical solution 31: A peptide containing a _VL region that is a functional variant of the antibody _VL region of technical solution 2.

技术方案32：如技术方案31所述的肽，其中肽的V_L区主要由与如SEQ ID No：2中所述序列的氨基酸同一性至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至 少约95％的序列组成。Technical scheme 32: the peptide as described in technical scheme 31, wherein the _VL district of peptide is mainly made up of at least about 50%, at least about 60%, at least about 70% with the amino acid identity of sequence as described in SEQ ID No:2 , at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% sequence composition.

技术方案33：含有作为技术方案2的抗体V_H区功能变异体的V_H 区的肽。Technical Solution 33: A peptide containing a _VH region that is a functional variant of the antibody _VH region of Technical Solution 2.

技术方案34：如技术方案33所述的肽，其中肽的V_H区主要由与如SEQ ID No：7中所述序列的氨基酸同一性至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％的序列组成。Technical scheme 34: the peptide as described in technical scheme 33, wherein the _VH region of peptide is mainly composed of at least about 50%, at least about 60%, at least about 70% of the amino acid identity of the sequence as described in SEQ ID No: 7 , at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% sequence composition.

技术方案35：至少含有一个作为技术方案2中抗体CDR的功能变异体的CDR的肽。Technical scheme 35: A peptide containing at least one CDR that is a functional variant of the antibody CDR in technical scheme 2.

技术方案36：如技术方案35所述的肽，其中至少一个肽的CDRs主要由与如SEQ IDNo：3、SEQ ID No：4、SEQ ID No：5、SEQ ID No：8、SEQ ID No：9或SEQ ID No：10中所述序列的氨基酸同一性至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％的序列组成。Technical scheme 36: The peptide as described in technical scheme 35, wherein the CDRs of at least one peptide are mainly composed of such as SEQ ID No: 3, SEQ ID No: 4, SEQ ID No: 5, SEQ ID No: 8, SEQ ID No: 9 or the amino acid identity of the sequence set forth in SEQ ID No: 10 is at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or At least about 95% sequence composition.

技术方案37：如技术方案31到36任何一项中所述的肽，其中肽具有至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％技术方案2中抗体的抗原决定簇结合特性。Technical scheme 37: The peptide as described in any one of technical scheme 31 to 36, wherein peptide has at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85% %, at least about 90%, or at least about 95% of the antigenic determinant binding properties of the antibody in technical scheme 2.

技术方案38：如技术方案31到36任何一项中所述的肽，其中肽具有至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％技术方案2中抗体的亲和性、强度或特异性。Technical scheme 38: The peptide as described in any one of technical scheme 31 to 36, wherein peptide has at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85% %, at least about 90% or at least about 95% of the affinity, strength or specificity of the antibody in technical scheme 2.

技术方案39：如技术方案12到38任何一项中所述的肽，该肽与人CD38特异地结合。Technical solution 39: The peptide according to any one of technical solutions 12 to 38, which specifically binds to human CD38.

技术方案40：如技术方案12到39任何一项中所述的肽，该肽与如技术方案2所述的抗体竞争结合CD38。Technical scheme 40: The peptide according to any one of technical schemes 12 to 39, which competes with the antibody according to technical scheme 2 for binding to CD38.

技术方案41：如技术方案40所述的肽，其中竞争作用是通过说明书的实施例8或9中所述的ELISA测定的。Technical Solution 41: The peptide according to Technical Solution 40, wherein the competition is determined by ELISA described in Example 8 or 9 of the description.

技术方案42：如技术方案7所述的肽，其中竞争作用是通过说明书的实施例7中所述的交叉抑制方法测定的。Technical solution 42: The peptide according to technical solution 7, wherein the competition is determined by the cross-inhibition method described in Example 7 of the specification.

技术方案43：如技术方案39所述的肽，该肽与CD38抗原决定簇特异结合，该抗原决定簇也与如技术方案2所述的抗体特异结合。Technical solution 43: The peptide according to technical solution 39, which specifically binds to the CD38 epitope, and the antigenic determinant also specifically binds to the antibody according to technical solution 2.

技术方案44：如技术方案39到43任何一项中所述的肽，其中肽与人CD38的结合亲和力大于如技术方案2所述的抗体。Technical scheme 44: The peptide according to any one of technical schemes 39 to 43, wherein the binding affinity of the peptide to human CD38 is greater than that of the antibody according to technical scheme 2.

技术方案45：如技术方案39到43任何一项中所述的肽，其中肽实质具有与如技术方案2所述的抗体相同的特异的CD38结合特性。Technical scheme 45: The peptide according to any one of technical schemes 39 to 43, wherein the peptide has substantially the same specific CD38-binding property as that of the antibody according to technical scheme 2.

技术方案46：如技术方案39到45任何一项中所述的肽，其中CD38结合肽实质不含有其它CD38结合肽。Technical solution 46: The peptide according to any one of technical solutions 39 to 45, wherein the CD38-binding peptide does not substantially contain other CD38-binding peptides.

技术方案47：与人CD38(SEQ ID No：31)结合的肽，它与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ IDNo：34)的结合程度不同于与其与人CD38(SEQ IDNo：31)的结合程度。Technical solution 47: A peptide that binds to human CD38 (SEQ ID No: 31) to a different extent from mutated human CD38 (SEQ ID No: 34) in which the 274-position serine residue is replaced by a phenylalanine residue in the degree of its binding to human CD38 (SEQ ID No: 31).

技术方案48：如技术方案47所述的肽，其中肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的50％。Technical scheme 48: The peptide according to technical scheme 47, wherein the EC ₅₀ of the peptide binding to mutated human CD38 (SEQ ID No: 34) in which the 274-position serine residue is replaced by a phenylalanine residue is less than the ₅₀ % of the EC50 of the peptide binding to human CD38 (SEQ ID No: 31).

技术方案49：如技术方案48所述的肽，其中肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的10％。Technical scheme 49: The peptide according to technical scheme 48, wherein the EC ₅₀ of the peptide binding to mutated human CD38 (SEQ ID No: 34) in which the 274-position serine residue is replaced by a phenylalanine residue is less than the 10% of the _EC50 of the peptide binding to human CD38 (SEQ ID No: 31).

技术方案50：如技术方案49所述的肽，其中肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的5％。Technical scheme 50: The peptide according to technical scheme 49, wherein the EC ₅₀ of the peptide binding to mutated human CD38 (SEQ ID No: 34) in which the 274-position serine residue is replaced by a phenylalanine residue is less than the 5% of the _EC50 of the peptide binding to human CD38 (SEQ ID No: 31).

技术方案51：如技术方案50所述的肽，其中肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的1％。Technical scheme 51: The peptide according to technical scheme 50, wherein the EC ₅₀ of the peptide binding to mutated human CD38 (SEQ ID No: 34) in which the 274-position serine residue is replaced by a phenylalanine residue is less than the 1% of the _EC50 of the peptide binding to human CD38 (SEQ ID No: 31).

技术方案52：与人CD38(SEQ ID No：31)结合的肽，它与其中272位谷氨酰胺残基被精氨酸残基替代的、突变的人CD38(SEQ IDNo：33)的结合程度不同于与其与人CD38(SEQ IDNo：31)的结合程度。Technical scheme 52: Peptide that binds to human CD38 (SEQ ID No: 31), its degree of binding to mutated human CD38 (SEQ ID No: 33) in which glutamine residue at position 272 is replaced by arginine residue Different from its binding degree to human CD38 (SEQ ID No: 31).

技术方案53：如技术方案52所述的肽，其中肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的50％。Technical scheme 53: The peptide according to technical scheme 52, wherein the EC ₅₀ of the peptide binding to mutated human CD38 (SEQ ID No: 34) in which the 274-position serine residue is replaced by a phenylalanine residue is less than the ₅₀ % of the EC50 of the peptide binding to human CD38 (SEQ ID No: 31).

技术方案54：如技术方案53所述的肽，其中肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的10％。Technical scheme 54: The peptide according to technical scheme 53, wherein the EC ₅₀ of the peptide binding to mutated human CD38 (SEQ ID No: 34) in which the 274-position serine residue is replaced by a phenylalanine residue is less than the 10% of the _EC50 of the peptide binding to human CD38 (SEQ ID No: 31).

技术方案55：技术方案47到51任何一项中的肽与其中272位谷氨酰胺残基被精氨酸残基替代的、突变的人CD38(SEQ ID No：33)的结合程度不同于与其与人CD38(SEQ IDNo：31)的结合程度。Technical scheme 55: The peptide of any one of technical schemes 47 to 51 binds to a mutant human CD38 (SEQ ID No: 33) in which the glutamine residue at position 272 is replaced by an arginine residue to a different degree than that of The degree of binding to human CD38 (SEQ ID No: 31).

技术方案56：如技术方案55所述的肽，其中肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的50％。Technical scheme 56: The peptide according to technical scheme 55, wherein the EC ₅₀ of the peptide binding to mutated human CD38 (SEQ ID No: 34) in which the 274-position serine residue is replaced by a phenylalanine residue is less than the ₅₀ % of the EC50 of the peptide binding to human CD38 (SEQ ID No: 31).

技术方案57：如技术方案56所述的肽，其中肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的10％。Technical scheme 57: The peptide according to technical scheme 56, wherein the EC ₅₀ of the peptide binding to mutated human CD38 (SEQ ID No: 34) in which the 274-position serine residue is replaced by a phenylalanine residue is less than the 10% of the _EC50 of the peptide binding to human CD38 (SEQ ID No: 31).

技术方案58：如技术方案47到57任何一项中所述的肽，其中所述肽肽与其中237位苏氨酸残基被丙氨酸残基替代的、突变的人CD38(SEQ ID No：32)结合，结合程度与其和人CD38(SEQ ID No：31)结合相同。Technical scheme 58: The peptide as described in any one of technical scheme 47 to 57, wherein said peptide peptide and wherein 237 threonine residues are replaced by alanine residues, mutated human CD38 (SEQ ID No :32) to the same extent as it binds to human CD38 (SEQ ID No: 31).

技术方案59：如技术方案58所述的肽，其中肽与突变的其中237位苏氨酸残基被丙氨酸残基替代的人CD38结合(SEQ ID No：32)的EC₅₀大于肽与人CD38(SEQ ID No：31)结合的EC₅₀的75％。Technical scheme 59: The peptide according to technical scheme 58, wherein the EC ₅₀ of the peptide binding to the mutated human CD38 (SEQ ID No: 32) in which the 237-threonine residue is replaced by an alanine residue is greater than that of the peptide and 75% of the _EC50 for human CD38 (SEQ ID No: 31 ) binding.

技术方案60：如技术方案59所述的肽，其中肽与突变的其中237位苏氨酸残基被丙氨酸残基替代的人CD38结合(SEQ ID No：32)的EC₅₀大于肽与人CD38(SEQ ID No：31)结合的EC₅₀的85％。Technical scheme 60: The peptide according to technical scheme 59, wherein the EC ₅₀ of the peptide binding to the mutated human CD38 (SEQ ID No: 32) in which the 237-threonine residue is replaced by an alanine residue is greater than that of the peptide and 85% of the _EC50 for human CD38 (SEQ ID No: 31 ) binding.

技术方案61：如技术方案60所述的肽，其中肽与突变的其中237位苏氨酸残基被丙氨酸残基替代的人CD38结合(SEQ ID No：32)的EC₅₀大于肽与人CD38(SEQ ID No：31)结合的EC₅₀的90％。Technical scheme 61: The peptide according to technical scheme 60, wherein the EC ₅₀ of the peptide binding to a mutated human CD38 (SEQ ID No: 32) in which the 237-threonine residue is replaced by an alanine residue is greater than that of the peptide and 90% of the _EC50 for human CD38 (SEQ ID No: 31 ) binding.

技术方案62：如技术方案6 1所述的肽，其中肽与突变的其中237位苏氨酸残基被丙氨酸残基替代的人CD38结合(SEQ ID No：32)的EC₅₀大于肽与人CD38(SEQ ID No：31)结合的EC₅₀的95％。Technical scheme 62: The peptide as described in technical scheme 61, wherein the EC ₅₀ of the peptide binding to the mutated human CD38 (SEQ ID No: 32) in which the threonine residue at position 237 is replaced by an alanine residue is greater than that of the peptide 95% of _EC50 for binding to human CD38 (SEQ ID No: 31).

技术方案63：与如技术方案4所述的抗体竞争结合CD38的肽。Technical scheme 63: A peptide that competes with the antibody described in technical scheme 4 for binding to CD38.

技术方案64：如技术方案63所述的肽，其中竞争作用是通过说明书的实施例8或9中所述的ELISA测定的。Technical solution 64: The peptide according to technical solution 63, wherein the competition is determined by ELISA described in Example 8 or 9 of the specification.

技术方案65：如技术方案63所述的肽，其中竞争作用是通过说明书的实施例7中所述的交叉抑制方法测定的。Technical Solution 65: The peptide according to Technical Solution 63, wherein the competition is determined by the cross-inhibition method described in Example 7 of the specification.

技术方案66：与CD38抗原决定簇特异结合的肽，该抗原决定簇也与如技术方案4所述的抗体特异结合。Technical scheme 66: A peptide that specifically binds to a CD38 epitope, and the antigenic determinant also specifically binds to the antibody described in technical scheme 4.

技术方案67：实质具有与如技术方案4所述的抗体相同的结合人CD38的特异结合特性的肽。Technical solution 67: A peptide having substantially the same specific binding property to human CD38 as the antibody described in technical solution 4.

技术方案68：含有主要由SEQ ID No：13组成的V_L CDR1的肽。Technical solution 68: A peptide containing V _L CDR1 mainly composed of SEQ ID No:13.

技术方案69：含有主要由SEQ ID No：14组成的V_L CDR2的肽。Technical scheme 69: A peptide containing _VL CDR2 mainly composed of SEQ ID No:14.

技术方案70：含有主要由SEQ ID No：15组成的V_L CDR3的肽。Technical solution 70: A peptide containing V _L CDR3 mainly composed of SEQ ID No: 15.

技术方案71：如技术方案70所述的肽，该肽也含有主要由SEQ IDNo：13组成的V_LCDR1。Technical solution 71: The peptide according to technical solution 70, which also contains V _L CDR1 mainly composed of SEQ ID No: 13.

技术方案72：如技术方案70所述的肽，该肽也含有主要由SEQ IDNo：14组成的V_LCDR2。Technical solution 72: The peptide according to technical solution 70, which also contains V _L CDR2 mainly composed of SEQ ID No: 14.

技术方案73：如技术方案72所述的肽，该肽也含有主要由SEQ IDNo：13组成的V_LCDR1。Technical solution 73: The peptide according to technical solution 72, which also contains V _L CDR1 mainly composed of SEQ ID No: 13.

技术方案74：含有主要由SEQ ID No：18组成的V_H CDR1的肽。Technical solution 74: A peptide containing _VH CDR1 mainly composed of SEQ ID No: 18.

技术方案75：含有主要由SEQ ID No：19组成的V_H CDR2的肽。Technical solution 75: A peptide containing _VH CDR2 mainly composed of SEQ ID No: 19.

技术方案76：含有主要由SEQ ID No：20组成的V_H CDR3的肽。Technical solution 76: A peptide containing _VH CDR3 mainly composed of SEQ ID No:20.

技术方案77：如技术方案76所述的肽，该肽也含有主要由SEQ IDNo：18组成的V_HCDR1。Technical scheme 77: The peptide according to technical scheme 76, which also contains _VH CDR1 mainly composed of SEQ ID No: 18.

技术方案78：如技术方案76所述的肽，该肽也含有主要由SEQ IDNo：19组成的V_HCDR2。Technical scheme 78: The peptide according to technical scheme 76, which also contains _VH CDR2 mainly composed of SEQ ID No: 19.

技术方案79：如技术方案78所述的肽，该肽也含有主要由SEQ IDNo：18组成的V_HCDR1。Technical scheme 79: The peptide according to technical scheme 78, which also contains _VH CDR1 mainly composed of SEQ ID No: 18.

技术方案80：肽，含有Technical scheme 80: peptide, containing

(a)含有3个V_L CDRs的第一个V_L区，它们彼此间各自主要由SEQ ID No：13、SEQ IDNo：14和SEQ ID No：15组成；及(a) a first _VL region comprising 3 _VL CDRs, each of which consists essentially of SEQ ID No: 13, SEQ ID No: 14 and SEQ ID No: 15; and

(b)含有3个V_H CDRs的第一个V_H区，它们彼此间各自主要由SEQ ID No：18、SEQ IDNo：19和SEQ ID No：20组成。(b) The first _VH region containing 3 _VH CDRs, which consist essentially of SEQ ID No: 18, SEQ ID No: 19 and SEQ ID No: 20 each other.

技术方案81：如技术方案80所述的肽，其中V_L区和V_H区位于肽的同一条链上。Technical solution 81: The peptide according to technical solution 80, wherein the _VL region and the _VH region are located on the same chain of the peptide.

技术方案82：如技术方案81所述的肽，其中V_L区和V_H区由柔性的连接物分隔开。Technical solution 82: The peptide according to technical solution 81, wherein the _VL region and the _VH region are separated by a flexible linker.

技术方案83：如技术方案80所述的肽，其中V_L区和V_H区位于肽的不同的链上。Technical solution 83: The peptide according to technical solution 80, wherein the _VL region and the _VH region are located on different chains of the peptide.

技术方案84：如技术方案83所述的肽，其中V_L区和V_H区位于作为免疫球蛋白折叠蛋白的肽的不同链上。Technical solution 84: The peptide according to technical solution 83, wherein the _VL region and the _VH region are located on different chains of the peptide which is an immunoglobulin folded protein.

技术方案85：如技术方案80到84任何一项中所述的肽，其中第一 个V_L区和第一个V_H区是定向的，从而使V_L区的3个CDRs和V_H区的3个CDRs协同作用来选择性地和/或特异性地与人CD38上的抗原决定簇结合。Technical scheme 85: The peptide as described in any one of technical scheme 80 to 84, wherein the first _VL district and the first _VH district are oriented, thereby make 3 CDRs of _VL district and _VH district The three CDRs of the CD38 work together to selectively and/or specifically bind to epitopes on human CD38.

技术方案86：如技术方案85所述的肽，其中肽含有第二个与第一个V_L区相同的V_L区和第二个与第一个V_H区相同的V_H区，其中第二个V_L区和第二个V_H区协同作用来选择性地和/或特异性地与人CD38上的抗原决定簇结合。Technical solution 86: The peptide of technical solution 85, wherein the peptide contains a second _VL region identical to the first _VL region and a second _VH region identical to the first _VH region, wherein the second The two _VL domains and the second _VH domain cooperate to selectively and/or specifically bind to antigenic determinants on human CD38.

技术方案87：含有作为技术方案4的抗体V_L区功能变异体的V_L 区的肽。Technical solution 87: A peptide containing a _VL region that is a functional variant of the antibody _VL region of technical solution 4.

技术方案88：如技术方案87所述的肽，其中肽的V_L区主要由与如SEQ ID No：12中所述序列的氨基酸同一性至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％的序列组成。Technical scheme 88: the peptide as described in technical scheme 87, wherein the _VL region of peptide is mainly composed of at least about 50%, at least about 60%, at least about 70% of the amino acid identity of the sequence as described in SEQ ID No: 12 , at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% sequence composition.

技术方案89：含有作为技术方案4的抗体V_H区功能变异体的V_H 区的肽。Technical solution 89: A peptide containing a _VH region that is a functional variant of the antibody _VH region of technical solution 4.

技术方案90：如技术方案89所述的肽，其中肽的V_H区主要由与如SEQ ID No：17中所述序列的氨基酸同一性至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％的序列组成。Technical scheme 90: the peptide as described in technical scheme 89, wherein the _VH region of peptide is mainly composed of at least about 50%, at least about 60%, at least about 70% of the amino acid identity of the sequence as described in SEQ ID No: 17 , at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% sequence composition.

技术方案91：至少含有一个作为技术方案4的抗体CDR的功能变异体的CDR的肽。Technical scheme 91: A peptide containing at least one CDR which is a functional variant of the antibody CDR of technical scheme 4.

技术方案92：如技术方案91所述的肽，其中至少一个肽的CDRs主要由与如SEQ IDNo：13、SEQ ID No：14、SEQ ID No：15、SEQ IDNo：18、SEQ ID No：19或SEQ ID No：20中所述序列的氨基酸同一性至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％的序列组成。Technical scheme 92: The peptide as described in technical scheme 91, wherein the CDRs of at least one peptide are mainly composed of such as SEQ ID No: 13, SEQ ID No: 14, SEQ ID No: 15, SEQ ID No: 18, SEQ ID No: 19 Or the amino acid identity of the sequence set forth in SEQ ID No: 20 is at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least Approximately 95% sequence composition.

技术方案93：如技术方案87到92任何一项中所述的肽，其中肽具有至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％技术方案4中抗体的抗原决定簇结合特性。Technical scheme 93: The peptide as described in any one of technical scheme 87 to 92, wherein peptide has at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85% %, at least about 90%, or at least about 95% of the antigenic determinant binding properties of the antibody in technical scheme 4.

技术方案94：如技术方案87到92任何一项中所述的肽，其中肽具有至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、 至少约85％、至少约90％或至少约95％技术方案4中抗体的亲和性、强度或特异性。Technical solution 94: The peptide as described in any one of technical solution 87 to 92, wherein the peptide has at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85% %, at least about 90% or at least about 95% of the affinity, strength or specificity of the antibody in technical scheme 4.

技术方案95：如技术方案68到94任何一项中所述的肽，该肽与人CD38特异地结合。Technical Item 95: The peptide according to any one of Technical Items 68 to 94, which specifically binds to human CD38.

技术方案96：如技术方案68到95任何一项中所述的肽，该肽与如技术方案4所述的抗体竞争结合CD38。Technical scheme 96: The peptide according to any one of technical schemes 68 to 95, which competes with the antibody according to technical scheme 4 for binding to CD38.

技术方案97：如技术方案96所述的肽，其中竞争作用是通过说明书的实施例8或9中所述的ELISA测定的。Technical scheme 97: The peptide according to technical scheme 96, wherein the competition is determined by ELISA described in Example 8 or 9 of the specification.

技术方案98：如技术方案96所述的肽，其中竞争作用是通过说明书的实施例7中所述的交叉抑制方法测定的。Technical Solution 98: The peptide according to Technical Solution 96, wherein the competition is determined by the cross-inhibition method described in Example 7 of the specification.

技术方案99：如技术方案95所述的肽，该肽与CD38抗原决定簇特异结合，该抗原决定簇也与如技术方案4所述的抗体特异结合。Technical scheme 99: The peptide according to technical scheme 95, which specifically binds to the CD38 epitope, and the antigenic determinant also specifically binds to the antibody according to technical scheme 4.

技术方案100：如技术方案95到99任何一项中所述的肽，其中肽与人CD38的结合亲和力大于如技术方案4所述的抗体。Technical scheme 100: the peptide as described in any one of technical schemes 95 to 99, wherein the binding affinity of the peptide to human CD38 is greater than that of the antibody as described in technical scheme 4.

技术方案101：如技术方案95到99任何一项中所述的肽，其中肽实质具有与如技术方案4所述的抗体相同的特异的CD38结合特性。Technical scheme 101: The peptide according to any one of technical schemes 95 to 99, wherein the peptide has substantially the same specific CD38-binding property as that of the antibody according to technical scheme 4.

技术方案102：如技术方案95到101任何一项中所述的肽，其中CD38结合肽实质不含有其它CD38结合肽。Technical solution 102: The peptide according to any one of technical solutions 95 to 101, wherein the CD38-binding peptide does not substantially contain other CD38-binding peptides.

技术方案103：如技术方案63到102任何一项中所述的肽，它与人CD38(SEQ ID No：31)结合，但不与突变的其中274位丝氨酸残基被苯丙氨酸残基替代的人CD38(SEQ ID No：34)如与其和人CD38(SEQID No：31)相同的结合程度进行结合。Technical solution 103: The peptide as described in any one of technical solutions 63 to 102, which binds to human CD38 (SEQ ID No: 31), but does not bind to the mutated 274th serine residue replaced by phenylalanine residue The substituted human CD38 (SEQ ID No: 34) binds to the same extent as it binds to human CD38 (SEQ ID No: 31).

技术方案104：如技术方案103所述的肽，其中肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的50％。Technical scheme 104: The peptide according to technical scheme 103, wherein the EC ₅₀ of the peptide binding to mutated human CD38 (SEQ ID No: 34) in which the 274-position serine residue is replaced by a phenylalanine residue is less than the ₅₀ % of the EC50 of the peptide binding to human CD38 (SEQ ID No: 31).

技术方案105：如技术方案104所述的肽，其中肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的10％。Technical scheme 105: The peptide according to technical scheme 104, wherein the EC ₅₀ of the peptide binding to mutated human CD38 (SEQ ID No: 34) in which the 274-position serine residue is replaced by a phenylalanine residue is less than the 10% of the _EC50 of the peptide binding to human CD38 (SEQ ID No: 31).

技术方案106：如技术方案105所述的肽，其中肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的5％。Technical scheme 106: The peptide according to technical scheme 105, wherein the EC ₅₀ of the peptide binding to mutated human CD38 (SEQ ID No: 34) in which the 274-position serine residue is replaced by a phenylalanine residue is less than the 5% of the _EC50 of the peptide binding to human CD38 (SEQ ID No: 31).

技术方案107：如技术方案106所述的肽，其中肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的1％。Technical scheme 107: The peptide according to technical scheme 106, wherein the EC ₅₀ of the peptide binding to mutated human CD38 (SEQ ID No: 34) in which the 274-position serine residue is replaced by a phenylalanine residue is less than the 1% of the _EC50 of the peptide binding to human CD38 (SEQ ID No: 31).

技术方案108：与人CD38(SEQ ID No：31)结合的肽，它与其中272位谷氨酰胺胺残基被精氨酸残基替代的、突变的人CD38(SEQ IDNo：33)的结合程度不同于与其与人CD38(SEQ ID No：31)的结合程度。Technical solution 108: Peptide binding to human CD38 (SEQ ID No: 31), which binds to mutated human CD38 (SEQ ID No: 33) in which glutamine residue at position 272 is replaced by arginine residue The extent differs from its binding to human CD38 (SEQ ID No: 31).

技术方案109：如技术方案108所述的肽，其中肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的50％。Technical scheme 109: The peptide according to technical scheme 108, wherein the EC ₅₀ of the peptide binding to mutated human CD38 (SEQ ID No: 34) in which the 274-position serine residue is replaced by a phenylalanine residue is less than the ₅₀ % of the EC50 of the peptide binding to human CD38 (SEQ ID No: 31).

技术方案110：如技术方案109所述的肽，其中肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的10％。Technical scheme 110: The peptide according to technical scheme 109, wherein the EC ₅₀ of the peptide binding to mutated human CD38 (SEQ ID No: 34) in which the 274-position serine residue is replaced by a phenylalanine residue is less than the 10% of the _EC50 of the peptide binding to human CD38 (SEQ ID No: 31).

技术方案111：技术方案103到107任何一项中的肽与其中272位谷氨酰胺残基被精氨酸残基替代的、突变的人CD38(SEQ ID No：33)的结合程度与其和人CD38(SEQ ID No：31)结合程度不同。Technical scheme 111: The degree of binding of the peptide in any one of technical schemes 103 to 107 to the mutated human CD38 (SEQ ID No: 33) in which the glutamine residue at position 272 is replaced by an arginine residue and human CD38 (SEQ ID No: 31) was bound to varying degrees.

技术方案112：如技术方案111所述的肽，其中肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的50％。Technical scheme 112: The peptide according to technical scheme 111, wherein the EC ₅₀ of the peptide binding to mutated human CD38 (SEQ ID No: 34) in which the 274-position serine residue is replaced by a phenylalanine residue is less than the ₅₀ % of the EC50 of the peptide binding to human CD38 (SEQ ID No: 31).

技术方案113：如技术方案112所述的肽，其中肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的10％。Technical scheme 113: The peptide according to technical scheme 112, wherein the EC ₅₀ of the peptide binding to mutated human CD38 (SEQ ID No: 34) in which the 274-position serine residue is replaced by a phenylalanine residue is less than the 10% of the _EC50 of the peptide binding to human CD38 (SEQ ID No: 31).

技术方案114：如技术方案103到113任何一项中所述的肽，其中所述肽与其中237位苏氨酸残基被丙氨酸残基替代的、突变的人CD38(SEQ ID No：32)结合，结合程度与其和人CD38(SEQ ID No：31)结合相同。Technical solution 114: The peptide as described in any one of technical solutions 103 to 113, wherein the peptide is mutated with human CD38 (SEQ ID No: 32) Binding, the degree of binding is the same as that of human CD38 (SEQ ID No: 31).

技术方案115：如技术方案114所述的肽，其中肽与突变的其中237位苏氨酸残基被丙氨酸残基替代的人CD38结合(SEQ ID No：32)的EC₅₀大于肽与人CD38(SEQ ID No：31)结合的EC₅₀的75％。Technical scheme 115: The peptide according to technical scheme 114, wherein the EC ₅₀ of the peptide binding to the mutated human CD38 (SEQ ID No: 32) in which the 237-threonine residue is replaced by an alanine residue is greater than that of the peptide and 75% of the _EC50 for human CD38 (SEQ ID No: 31 ) binding.

技术方案116：如技术方案115所述的肽，其中肽与突变的其中237位苏氨酸残基被丙氨酸残基替代的人CD38结合(SEQ ID No：32)的EC₅₀大于肽与人CD38(SEQ ID No：31)结合的EC₅₀的85％。Technical scheme 116: The peptide according to technical scheme 115, wherein the EC ₅₀ of the peptide binding to the mutated human CD38 (SEQ ID No: 32) in which the 237-threonine residue is replaced by an alanine residue is greater than that of the peptide and 85% of the _EC50 for human CD38 (SEQ ID No: 31 ) binding.

技术方案117：如技术方案116所述的肽，其中肽与突变的其中237位苏氨酸残基被丙氨酸残基替代的人CD38结合(SEQ ID No：32)的EC₅₀大于肽与人CD38(SEQ ID No：31)结合的EC₅₀的90％。Technical scheme 117: The peptide according to technical scheme 116, wherein the EC ₅₀ of the peptide binding to the mutated human CD38 (SEQ ID No: 32) in which the 237-threonine residue is replaced by an alanine residue is greater than that of the peptide and 90% of the _EC50 for human CD38 (SEQ ID No: 31 ) binding.

技术方案118：如技术方案117所述的肽，其中肽与突变的其中237位苏氨酸残基被丙氨酸残基替代的人CD38结合(SEQ ID No：32)的EC₅₀大于肽与人CD38(SEQ ID No：31)结合的EC₅₀的95％。Technical scheme 118: The peptide according to technical scheme 117, wherein the EC ₅₀ of the peptide binding to a mutated human CD38 (SEQ ID No: 32) in which the 237-threonine residue is replaced by an alanine residue is greater than that of the peptide and 95% of the _EC50 for human CD38 (SEQ ID No: 31 ) binding.

技术方案119：与如技术方案6所述的抗体竞争结合CD38的肽。Technical scheme 119: A peptide that competes with the antibody described in technical scheme 6 for binding to CD38.

技术方案120：如技术方案119所述的肽，其中竞争作用是通过说明书的实施例8或9中所述的ELISA测定的。Technical solution 120: The peptide according to technical solution 119, wherein the competition is determined by ELISA described in Example 8 or 9 of the description.

技术方案121：如技术方案119所述的肽，其中竞争作用是通过说明书的实施例7中所述的交叉抑制方法测定的。Technical solution 121: The peptide according to technical solution 119, wherein the competition is determined by the cross-inhibition method described in Example 7 of the specification.

技术方案122：与CD38抗原决定簇特异结合的肽，该抗原决定簇也与如技术方案6所述的抗体特异结合。Technical scheme 122: a peptide that specifically binds to a CD38 epitope, and the antigenic determinant also specifically binds to the antibody described in technical scheme 6.

技术方案123：实质具有与如技术方案6所述的抗体相同的结合人CD38的特异结合特性的肽。Technical solution 123: A peptide having substantially the same specific binding property to human CD38 as the antibody described in technical solution 6.

技术方案124：含有主要由SEQ ID No：23组成的V_L CDR1的肽。Technical solution 124: A peptide containing V _L CDR1 mainly composed of SEQ ID No:23.

技术方案125：含有主要由SEQ ID No：24组成的V_L CDR2的肽。Technical solution 125: A peptide containing V _L CDR2 mainly composed of SEQ ID No:24.

技术方案126：含有主要由SEQ ID No：25组成的V_L CDR3的肽。Technical solution 126: A peptide containing _VL CDR3 mainly composed of SEQ ID No:25.

技术方案127：如技术方案126所述的肽，该肽也含有主要由SEQ IDNo：23组成的V_LCDR1。Technical scheme 127: The peptide according to technical scheme 126, which also contains V _L CDR1 mainly composed of SEQ ID No: 23.

技术方案128：如技术方案126所述的肽，该肽也含有主要由SEQ IDNo：24组成的V_LCDR2。Technical solution 128: The peptide according to technical solution 126, which also contains V _L CDR2 mainly composed of SEQ ID No: 24.

技术方案129：如技术方案128所述的肽，该肽也含有主要由SEQ IDNo：23组成的V_LCDR1。Technical scheme 129: The peptide according to technical scheme 128, which also contains V _L CDR1 mainly composed of SEQ ID No: 23.

技术方案130：含有主要由SEQ ID No：28组成的V_H CDR1的肽。Technical solution 130: A peptide containing _VH CDR1 mainly composed of SEQ ID No:28.

技术方案131：含有主要由SEQ ID No：29组成的V_H CDR2的肽。Technical solution 131: A peptide containing _VH CDR2 mainly composed of SEQ ID No:29.

技术方案132：含有主要由SEQ ID No：30组成的V_H CDR3的肽。Technical solution 132: A peptide containing _VH CDR3 mainly composed of SEQ ID No:30.

技术方案133：如技术方案132所述的肽，该肽也含有主要由SEQ IDNo：28组成的V_HCDR1。Technical scheme 133: The peptide according to technical scheme 132, which also contains _VH CDR1 mainly composed of SEQ ID No: 28.

技术方案134：如技术方案132所述的肽，该肽也含有主要由SEQ IDNo：29组成的V_HCDR2。Technical solution 134: The peptide according to technical solution 132, which also contains _VH CDR2 mainly composed of SEQ ID No: 29.

技术方案135：如技术方案134所述的肽，该肽也含有主要由SEQ IDNo：28组成的V_HCDR1。Technical scheme 135: The peptide according to technical scheme 134, which also contains _VH CDR1 mainly composed of SEQ ID No: 28.

技术方案136：肽，含有Technical scheme 136: peptide, containing

(a)含有3个V_L CDRs的第一个V_L区，它们彼此间各自主要由SEQ ID No：23、SEQ IDNo：24和SEQ ID No：25组成；及(a) a first _VL region comprising 3 _VL CDRs, each of which consists essentially of SEQ ID No: 23, SEQ ID No: 24 and SEQ ID No: 25; and

(b)含有3个V_H CDRs的第一个V_H区，它们彼此间各自主要由SEQ ID No：28、SEQ IDNo：29和SEQ ID No：30组成。(b) A first _VH region containing 3 _VH CDRs, which consist essentially of SEQ ID No: 28, SEQ ID No: 29 and SEQ ID No: 30 each other.

技术方案137：如技术方案136所述的肽，其中V_L区和V_H区位于肽的同一条链上。Technical solution 137: The peptide according to technical solution 136, wherein the _VL region and the _VH region are located on the same chain of the peptide.

技术方案138：如技术方案137所述的肽，其中V_L区和V_H区由柔性的连接物分隔开。Technical solution 138: The peptide according to technical solution 137, wherein the _VL region and the _VH region are separated by a flexible linker.

技术方案139：如技术方案136所述的肽，其中V_L区和V_H区位于肽的不同的链上。Technical solution 139: The peptide according to technical solution 136, wherein the _VL region and the _VH region are located on different chains of the peptide.

技术方案140：如技术方案139所述的肽，其中V_L区和V_H区位于作为免疫球蛋白折叠蛋白的肽的不同链上。Technical solution 140: The peptide according to technical solution 139, wherein the _VL region and the _VH region are located on different chains of the peptide which is an immunoglobulin folded protein.

技术方案141：如技术方案136到140任何一项中所述的肽，其中第一个V_L区和第一个V_H区是定向的，从而使V_L区的3个CDRs和V_H 区的3个CDRs协同作用来选择性地和/或特异性地与人CD38上的抗原决定簇结合。Technical scheme 141: The peptide as described in any one of technical scheme 136 to 140, wherein the first _VL district and the first _VH district are oriented, thereby make 3 CDRs of _VL district and _VH district The three CDRs of the CD38 work together to selectively and/or specifically bind to epitopes on human CD38.

技术方案142：如技术方案141所述的肽，其中肽含有第二个与第一个V_L区相同的V_L区和第二个与第一个V_H区相同的V_H区，其中第二个V_L区和第二个V_H区协同作用来选择性地和/或特异性地与人CD38上的抗原决定簇结合。Technical scheme 142: The peptide of technical scheme 141, wherein the peptide contains a second _VL region identical to the first _VL region and a second _VH region identical to the first _VH region, wherein the second The two _VL domains and the second _VH domain cooperate to selectively and/or specifically bind to antigenic determinants on human CD38.

技术方案143：含有作为技术方案6的抗体V_L区功能变异体的V_L 区的肽。Technical solution 143: A peptide containing a _VL region that is a functional variant of the antibody _VL region of technical solution 6.

技术方案144：如技术方案143所述的肽，其中肽的V_L区主要由与如SEQ ID No：22中所述序列的氨基酸同一性至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％的序列组成。Technical scheme 144: the peptide as described in technical scheme 143, wherein the V _L region of peptide is mainly composed of at least about 50%, at least about 60%, at least about 70% of the amino acid identity of the sequence as described in SEQ ID No:22 , at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% sequence composition.

技术方案145：含有作为技术方案6的抗体V_H区功能变异体的V_H 区的肽。Technical Solution 145: A peptide containing a _VH region that is a functional variant of the antibody _VH region of Technical Solution 6.

技术方案146：如技术方案145所述的肽，其中肽的V_H区主要由与如SEQ ID No：27中所述序列的氨基酸同一性至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％的序列组成。Technical scheme 146: the peptide as described in technical scheme 145, wherein the _VH region of peptide is mainly composed of at least about 50%, at least about 60%, at least about 70% of the amino acid identity of the sequence as described in SEQ ID No:27 , at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% sequence composition.

技术方案147：至少含有一个作为技术方案6的抗体CDR的功能变异体的CDR的肽。Technical scheme 147: A peptide containing at least one CDR that is a functional variant of the antibody CDR of technical scheme 6.

技术方案148：如技术方案147所述的肽，其中至少一个肽的CDRs主要由与如SEQID No：23、SEQ ID No：24、SEQ ID No：25、SEQ IDNo：28、SEQ ID No：29或SEQ ID No：30中所述序列的氨基酸同一性至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％的序列组成。Technical scheme 148: The peptide as described in technical scheme 147, wherein the CDRs of at least one peptide are mainly composed of such as SEQ ID No: 23, SEQ ID No: 24, SEQ ID No: 25, SEQ ID No: 28, SEQ ID No: 29 Or the amino acid identity of the sequence set forth in SEQ ID No: 30 is at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least Approximately 95% sequence composition.

技术方案149：如技术方案143到148任何一项中所述的肽，其中肽具有至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％技术方案6中抗体的抗原决定簇结合特性。Technical solution 149: The peptide as described in any one of technical solution 143 to 148, wherein the peptide has at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85% %, at least about 90%, or at least about 95% of the antigenic determinant binding properties of the antibody in technical scheme 6.

技术方案150：如技术方案143到148任何一项中所述的肽，其中肽具有至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％或至少约95％技术方案6中抗体的亲和性、强度或特异性。Technical scheme 150: the peptide as described in any one of technical scheme 143 to 148, wherein peptide has at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85% %, at least about 90% or at least about 95% of the affinity, strength or specificity of the antibody in technical scheme 6.

技术方案151：如技术方案124到150任何一项中所述的肽，该肽与人CD38特异地结合。Technical solution 151: The peptide according to any one of technical solutions 124 to 150, which specifically binds to human CD38.

技术方案152：如技术方案124到151任何一项中所述的肽，该肽与如技术方案6所述的抗体竞争结合CD38。Technical scheme 152: The peptide according to any one of technical schemes 124 to 151, which competes with the antibody according to technical scheme 6 for binding to CD38.

技术方案153：如技术方案152所述的肽，其中竞争作用是通过说明书的实施例8或9中所述的ELISA测定的。Technical solution 153: The peptide according to technical solution 152, wherein the competition is determined by the ELISA described in Example 8 or 9 of the description.

技术方案154：如技术方案152所述的肽，其中竞争作用是通过说明书的实施例7中所述的交叉抑制方法测定的。Technical solution 154: The peptide according to technical solution 152, wherein the competition is determined by the cross-inhibition method described in Example 7 of the specification.

技术方案155：如技术方案151所述的肽，该肽与CD38抗原决定簇特异结合，该抗原决定簇也与如技术方案6所述的抗体特异结合。Technical scheme 155: The peptide according to technical scheme 151, which specifically binds to the CD38 epitope, and the antigenic determinant also specifically binds to the antibody according to technical scheme 6.

技术方案156：如技术方案151到155任何一项中所述的肽，其中肽与人CD38的结合亲和力大于如技术方案6所述的抗体。Technical scheme 156: The peptide as described in any one of technical schemes 151 to 155, wherein the binding affinity of the peptide to human CD38 is greater than that of the antibody described in technical scheme 6.

技术方案157：如技术方案151到155任何一项中所述的肽，其中肽实质具有与如技术方案6所述的抗体相同的特异的CD38结合特性。Technical scheme 157: The peptide according to any one of technical schemes 151 to 155, wherein the peptide has substantially the same specific CD38-binding property as that of the antibody according to technical scheme 6.

技术方案158：如技术方案151到157任何一项中所述的肽，其中CD38结合肽实质不含有其它CD38结合肽。Technical solution 158: The peptide according to any one of technical solutions 151 to 157, wherein the CD38-binding peptide does not substantially contain other CD38-binding peptides.

技术方案159：如技术方案1到158任何一项中所述的肽，其中肽 不是CD38的拮抗物。Technical solution 159: The peptide according to any one of technical solutions 1 to 158, wherein the peptide is not an antagonist of CD38.

技术方案160：如技术方案1到159任何一项中所述的肽，其中肽不诱导外周血单核细胞的显著增殖。Technical solution 160: The peptide according to any one of technical solutions 1 to 159, wherein the peptide does not induce significant proliferation of peripheral blood mononuclear cells.

技术方案161：如技术方案1到160任何一项中所述的肽，其中肽不诱导人单核细胞或外周血单核细胞显著释放IL-6。Technical solution 161: The peptide according to any one of technical solutions 1 to 160, wherein the peptide does not induce significant release of IL-6 from human monocytes or peripheral blood mononuclear cells.

技术方案162：如技术方案1到161任何一项中所述的肽，其中肽不诱导人T细胞或外周血单核细胞释放可检测的IFN-γ。Technical solution 162: The peptide according to any one of technical solutions 1 to 161, wherein the peptide does not induce human T cells or peripheral blood mononuclear cells to release detectable IFN-γ.

技术方案163：如技术方案1到162任何一项中所述的肽，其中肽是抗体。Technical scheme 163: The peptide as described in any one of technical schemes 1 to 162, wherein the peptide is an antibody.

技术方案164：如技术方案1到6或163任何一项中所述的抗体，其中抗体是人抗体。Technical scheme 164: The antibody according to any one of technical schemes 1 to 6 or 163, wherein the antibody is a human antibody.

技术方案165：如技术方案1到6或163任何一项中所述的抗体，其中抗体是人源化抗体。Technical solution 165: The antibody according to any one of technical solutions 1 to 6 or 163, wherein the antibody is a humanized antibody.

技术方案166：如技术方案1到6或163任何一项中所述的抗体，其中抗体是嵌合抗体。Technical scheme 166: The antibody according to any one of technical schemes 1 to 6 or 163, wherein the antibody is a chimeric antibody.

技术方案167：如技术方案1到6或163任何一项中所述的抗体，其中抗体是单克隆抗体。Technical scheme 167: The antibody according to any one of technical schemes 1 to 6 or 163, wherein the antibody is a monoclonal antibody.

技术方案168：如技术方案1到6或163任何一项中所述的抗体，其特征是它是IgG1、IgG2、IgG3、IgG4、IgD、IgA、IgE或IgM抗体。Technical solution 168: The antibody according to any one of technical solutions 1 to 6 or 163, characterized in that it is an IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE or IgM antibody.

技术方案169：如技术方案168所述的抗体，其特征是它是IgG1抗体。Technical solution 169: The antibody according to technical solution 168, which is characterized in that it is an IgG1 antibody.

技术方案170：如技术方案169所述的抗体，其特征是它是IgG1，κ抗体。Technical solution 170: The antibody according to technical solution 169, characterized in that it is an IgG1, κ antibody.

技术方案171：如技术方案168所述的抗体，其特征是它是IgM抗体。Technical solution 171: The antibody according to technical solution 168, which is characterized in that it is an IgM antibody.

技术方案172：如技术方案171所述的抗体，其中抗体是IgM1κ抗体。Technical solution 172: The antibody according to technical solution 171, wherein the antibody is an IgM1κ antibody.

技术方案173：如技术方案2到172任何一项中所述的肽，其中肽在真核细胞中是糖基化的。Technical item 173: The peptide according to any one of technical items 2 to 172, wherein the peptide is glycosylated in eukaryotic cells.

技术方案174：如技术方案1到6或163任何一项中所述的抗体，它是抗体片段或单链抗体。Technical scheme 174: The antibody according to any one of technical schemes 1 to 6 or 163, which is an antibody fragment or a single-chain antibody.

技术方案175：如技术方案1到174任何一项中所述的肽或抗体， 进一步含有用于连接放射性同位素的螯合连接物。Technical scheme 175: The peptide or antibody as described in any one of technical schemes 1 to 174, further comprising a chelate linker for linking a radioisotope.

技术方案176：如技术方案1到175任何一项中所述的肽，它实质是分离的形式。Technical scheme 176: The peptide according to any one of technical schemes 1 to 175, which is substantially in an isolated form.

技术方案177：编码如技术方案1到175任何一项中所述的肽的分离的核酸。Technical scheme 177: An isolated nucleic acid encoding the peptide described in any one of technical schemes 1 to 175.

技术方案178：含有编码如技术方案1到175任何一项中所述的肽的核酸的表达载体。Technical solution 178: An expression vector containing a nucleic acid encoding the peptide described in any one of technical solutions 1 to 175.

技术方案179：含有SEQ ID No：1中的V_L核苷酸序列、SEQ ID No：6中的V_H核苷酸序列或SEQ ID No：1中的V_L核苷酸序列和SEQ IDNo：6中的V_H核苷酸序列的表达载体。Technical solution 179: containing the V _L nucleotide sequence in SEQ ID No: 1, the V _H nucleotide sequence in SEQ ID No: 6 or the V _L nucleotide sequence in SEQ ID No: 1 and SEQ ID No: The expression vector of the _VH nucleotide sequence in 6.

技术方案180：含有SEQ ID No：11中的V_L核苷酸序列、SEQ IDNo：16中的V_H核苷酸序列或SEQ ID No：11中的V_L核苷酸序列和SEQID No：16中的V_H核苷酸序列的表达载体。Technical solution 180: containing the V _L nucleotide sequence in SEQ ID No: 11, the V _H nucleotide sequence in SEQ ID No: 16 or the V _L nucleotide sequence in SEQ ID No: 11 and SEQ ID No: 16 The expression vector of the _VH nucleotide sequence in.

技术方案181：如技术方案179或技术方案180所述的表达载体，进一步包括编码人抗体的轻链、重链或轻链和重链的恒定区的核苷酸序列。Technical scheme 181: The expression vector as described in technical scheme 179 or technical scheme 180, further comprising a nucleotide sequence encoding the constant region of the light chain, the heavy chain, or the light chain and the heavy chain of a human antibody.

技术方案182：如技术方案181所述的表达载体，其中编码人抗体的轻链、重链或轻链和重链的恒定区的核苷酸序列编码IgG1抗体。Technical solution 182: The expression vector as described in technical solution 181, wherein the nucleotide sequence encoding the constant region of the light chain, the heavy chain or the light chain and the heavy chain of a human antibody encodes an IgG1 antibody.

技术方案183：可产生由人轻链和人重链核酸编码的人单克隆抗CD38抗体的杂交瘤，这些序列包括如SEQ ID No：1中的可变轻链区的核苷酸序列或其保守序列改变，及如SEQ ID No：6中的可变重链区的核苷酸序列或其保守序列改变。Technical solution 183: A hybridoma that can produce human monoclonal anti-CD38 antibodies encoded by human light chain and human heavy chain nucleic acids, these sequences include the nucleotide sequence of the variable light chain region in SEQ ID No: 1 or its Conservative sequence changes, and the nucleotide sequence of the variable heavy chain region as in SEQ ID No: 6 or its conservative sequence changes.

技术方案184：如技术方案183所述的杂交瘤，其中人轻链核酸含有如SEQ ID No：1所示的核苷酸序列，人重链核酸含有如SEQ ID No：6所示的核苷酸序列。Technical solution 184: The hybridoma as described in technical solution 183, wherein the human light chain nucleic acid contains the nucleotide sequence shown in SEQ ID No: 1, and the human heavy chain nucleic acid contains the nucleotide sequence shown in SEQ ID No: 6 acid sequence.

技术方案185：可产生由人轻链和人重链核酸编码的人单克隆抗CD38抗体的杂交瘤，这些序列包括如SEQ ID No：2中的可变轻链区的核苷酸序列或其保守序列改变，及如SEQ ID No：7中的可变重链区的核苷酸序列或其保守序列改变。Technical solution 185: A hybridoma that can produce human monoclonal anti-CD38 antibodies encoded by human light chain and human heavy chain nucleic acids, these sequences include the nucleotide sequence of the variable light chain region in SEQ ID No: 2 or its Conservative sequence changes, and the nucleotide sequence of the variable heavy chain region as in SEQ ID No: 7 or its conservative sequence changes.

技术方案186：如技术方案185所述的杂交瘤，其中人轻链核酸含有如SEQ ID No：2所示的核苷酸序列，人重链核酸含有如SEQ ID No：7所示的核苷酸序列。Technical solution 186: The hybridoma as described in technical solution 185, wherein the human light chain nucleic acid contains the nucleotide sequence shown in SEQ ID No: 2, and the human heavy chain nucleic acid contains the nucleotide sequence shown in SEQ ID No: 7 acid sequence.

技术方案187：可产生由如SEQ ID No：1中的人轻链可变区的核酸或其保守序列改变及如SEQ ID No：6中的人重链核酸或其保守序列改变编码的人单克隆抗CD38抗体的转染瘤。Technical solution 187: Human mononucleic acid encoded by the nucleic acid of the human light chain variable region as in SEQ ID No: 1 or its conservative sequence change and the human heavy chain nucleic acid as in SEQ ID No: 6 or its conservative sequence change can be produced Cloning of transfectomas with anti-CD38 antibodies.

技术方案188：如技术方案187所述的转染瘤，其中人单克隆抗CD3 8抗体由如SEQID No：1中的人轻链可变区核酸和如SEQ ID No：6中的人重链核酸编码。Technical scheme 188: the transfectoma as described in technical scheme 187, wherein the human monoclonal anti-CD3 8 antibody is made of the human light chain variable region nucleic acid as in SEQ ID No: 1 and the human heavy chain as in SEQ ID No: 6 nucleic acid code.

技术方案189：可产生具有人轻链和重链可变区的人单克隆抗CD38抗体的转染瘤，这些可变区包括如SEQ ID No：2中的人轻链可变区的氨基酸序列或其保守序列改变，及如SEQ ID No：7中的人重链可变区的氨基酸序列或其保守序列改变。Technical solution 189: Transfectoma capable of producing human monoclonal anti-CD38 antibody with human light chain and heavy chain variable regions, these variable regions include the amino acid sequence of human light chain variable region as in SEQ ID No: 2 or its conservative sequence change, and the amino acid sequence of the human heavy chain variable region in SEQ ID No: 7 or its conservative sequence change.

技术方案190：如技术方案189所述的转染瘤，人轻链含有如SEQ IDNo：2中的人轻链可变区氨基酸序列，人重链含有如SEQ ID No：7中的人重链可变区氨基酸序列。Technical solution 190: the transfectoma as described in technical solution 189, the human light chain contains the amino acid sequence of the human light chain variable region as in SEQ ID No: 2, and the human heavy chain contains the human heavy chain as in SEQ ID No: 7 Variable region amino acid sequence.

技术方案191：可产生由人轻链和人重链核酸编码的人单克隆抗CD38抗体的杂交瘤，这些序列包括如SEQ ID No：11中的可变轻链区的核苷酸序列或其保守序列改变，及如SEQ ID No：16中的可变重链区的核苷酸序列或其保守序列改变。Technical solution 191: A hybridoma that can produce human monoclonal anti-CD38 antibodies encoded by human light chain and human heavy chain nucleic acids, these sequences include the nucleotide sequence of the variable light chain region in SEQ ID No: 11 or its Conservative sequence changes, and the nucleotide sequence of the variable heavy chain region as in SEQ ID No: 16 or its conservative sequence changes.

技术方案192：如技术方案191所述的杂交瘤，其中人轻链核酸含有如SEQ ID No：11所示的核苷酸序列，人重链核酸含有如SEQ ID No：16所示的核苷酸序列。Technical solution 192: The hybridoma as described in technical solution 191, wherein the human light chain nucleic acid contains the nucleotide sequence shown in SEQ ID No: 11, and the human heavy chain nucleic acid contains the nucleoside shown in SEQ ID No: 16 acid sequence.

技术方案193：可产生具有人重链和轻链可变区的人单克隆抗CD38抗体的杂交瘤，这些可变区包括如SEQ ID No：12中的人轻链可变区的氨基酸序列或其保守序列改变，及如SEQ ID No：17中的人重链可变区的氨基酸序列或其保守序列改变。Technical solution 193: A hybridoma capable of producing a human monoclonal anti-CD38 antibody having human heavy and light chain variable regions, these variable regions include the amino acid sequence of the human light chain variable region in SEQ ID No: 12 or Its conservative sequence changes, and the amino acid sequence of the human heavy chain variable region as in SEQ ID No: 17 or its conservative sequence changes.

技术方案194：如技术方案193所述的杂交瘤，其中人轻链可变区含有如SEQ IDNo：12中的氨基酸序列，人重链可变区含有如SEQ IDNo：17中的氨基酸序列。Technical solution 194: The hybridoma according to technical solution 193, wherein the human light chain variable region contains the amino acid sequence of SEQ ID No: 12, and the human heavy chain variable region contains the amino acid sequence of SEQ ID No: 17.

技术方案195：可产生由如SEQ ID No：11中的人轻链可变区的核酸或其保守序列改变及如SEQ ID No：16中的人重链核酸或其保守序列改变编码的人单克隆抗CD38抗体的转染瘤。Technical solution 195: It can produce human mononucleic acid encoded by the nucleic acid of the human light chain variable region as in SEQ ID No: 11 or its conservative sequence change and the human heavy chain nucleic acid as in SEQ ID No: 16 or its conservative sequence change Cloning of transfectomas with anti-CD38 antibodies.

技术方案196：如技术方案195所述的杂交瘤，其中人单克隆抗CD38抗体由如SEQID No：11中的人轻链可变区核酸和如SEQ ID No： 16中的人重链核酸编码。Technical scheme 196: The hybridoma as described in technical scheme 195, wherein the human monoclonal anti-CD38 antibody is encoded by the human light chain variable region nucleic acid as in SEQ ID No: 11 and the human heavy chain nucleic acid as in SEQ ID No: 16 .

技术方案197：可产生具有人轻链和重链可变区的人单克隆抗CD38抗体的转染瘤，这些可变区包括如SEQ ID No：12中的人轻链可变区的氨基酸序列或其保守序列改变，及如SEQ ID No：17中的人重链可变区的氨基酸序列或其保守序列改变。Technical scheme 197: Transfectoma capable of producing human monoclonal anti-CD38 antibody with human light chain and heavy chain variable regions, these variable regions include the amino acid sequence of human light chain variable region as in SEQ ID No: 12 or its conservative sequence change, and the amino acid sequence of the human heavy chain variable region in SEQ ID No: 17 or its conservative sequence change.

技术方案198：如技术方案197所述的杂交瘤，其中人轻链含有如SEQ ID No：12中的人轻链可变区氨基酸序列，人重链含有如SEQ IDNo：17中的人重链可变区氨基酸序列。Technical solution 198: The hybridoma as described in technical solution 197, wherein the human light chain contains the amino acid sequence of the human light chain variable region as in SEQ ID No: 12, and the human heavy chain contains the human heavy chain as in SEQ ID No: 17 Variable region amino acid sequence.

技术方案199：可产生如技术方案1到175任何一项中所述的肽的真核或原核宿主细胞。Technical scheme 199: A eukaryotic or prokaryotic host cell capable of producing the peptide described in any one of technical schemes 1 to 175.

技术方案200：含有如技术方案178所述的表达载体的真核或原核宿主细胞。Technical scheme 200: a eukaryotic or prokaryotic host cell containing the expression vector as described in technical scheme 178.

技术方案201：含有编码人重链和人轻链的核酸的转基因非人动物或植物，其中动物或植物产生可检测量的如技术方案1到175任何一项中所述的肽。Technical solution 201: A transgenic non-human animal or plant containing nucleic acid encoding a human heavy chain and a human light chain, wherein the animal or plant produces a detectable amount of the peptide described in any one of technical solutions 1 to 175.

技术方案202：含有与细胞毒药剂、放射性同位素或药物连接的如技术方案1到174任何一项中所述的肽的免疫连接物。Technical scheme 202: The immune linker comprising the peptide described in any one of technical schemes 1 to 174 linked to cytotoxic agents, radioisotopes or drugs.

技术方案203：含有与细胞毒药剂、放射性同位素或药物连接的如技术方案1到55或技术方案171到174任何一项中所述的肽的免疫连接物，其中肽是单价的与细胞毒素药剂、放射性同位素或药物连接的IgM抗体。Technical scheme 203: The immune conjugate containing the peptide described in any one of technical schemes 1 to 55 or technical schemes 171 to 174 linked to cytotoxic agents, radioisotopes or drugs, wherein the peptide is monovalent and cytotoxic agents , radioisotope or drug-linked IgM antibodies.

技术方案204：含如技术方案1到175任何一项中所述的肽并与人效应细胞有结合特异性的双特异性或多特异性分子。Technical solution 204: a bispecific or multispecific molecule comprising the peptide described in any one of technical solutions 1 to 175 and having binding specificity to human effector cells.

技术方案205：含如技术方案1到175任何一项中所述的肽并与CD3、CD4、CD138、IL-15R、膜结合或受体结合的TNF-α、人Fc受体、或膜结合或受体结合的IL-15有结合特异性的双特异性或多特异性分子。Technical scheme 205: TNF-α, human Fc receptor, or membrane-bound TNF-alpha, human Fc receptor, or membrane-bound or receptor-bound CD3, CD4, CD138, IL-15R, containing the peptide described in any one of technical schemes 1 to 175 Or bispecific or multispecific molecules with binding specificity for receptor-bound IL-15.

技术方案206：含如技术方案1到176任何一项中所述的肽或如技术方案202或205所述的免疫连接物及药学上可接受的载体的药物组合物。Technical scheme 206: a pharmaceutical composition containing the peptide as described in any one of technical schemes 1 to 176 or the immune linker as described in technical scheme 202 or 205 and a pharmaceutically acceptable carrier.

技术方案207：如技术方案206所述的药物组合物含有一种或多种其它的治疗性药剂。Technical scheme 207: The pharmaceutical composition as described in technical scheme 206 contains one or more other therapeutic agents.

技术方案208：一种抑制表达CD38的细胞的生长和/或增殖的方法，包括给药如技术方案1到176任何一项中所述的肽、如技术方案202或205所述的免疫连接物、如技术方案206或207所述的药物组合物，或如技术方案178到182任何一项中所述的表达载体，从而抑制细胞的生长和/或增殖。Technical scheme 208: A method for inhibiting the growth and/or proliferation of cells expressing CD38, comprising administering the peptide as described in any one of technical schemes 1 to 176, and the immune linker as described in technical scheme 202 or 205 , the pharmaceutical composition as described in technical scheme 206 or 207, or the expression vector as described in any one of technical schemes 178 to 182, thereby inhibiting the growth and/or proliferation of cells.

技术方案209：一种治疗患者由表达CD38的细胞参与的疾病或症状的方法，该方法包括对所需要治疗的患者给药如技术方案1到176任何一项中所述的肽、如技术方案202或205所述的免疫连接物、如技术方案206或207所述的药物组合物，或如技术方案178到182任何一项中所述的表达载体。Technical scheme 209: A method for treating a disease or symptom in a patient in which CD38-expressing cells are involved, the method comprising administering the peptide described in any one of technical schemes 1 to 176, such as the technical scheme The immune linker described in 202 or 205, the pharmaceutical composition as described in technical scheme 206 or 207, or the expression vector as described in any one of technical schemes 178 to 182.

技术方案210：一种抑制患者由表达CD38的细胞参与的疾病或症状的方法，该方法包括对所需要治疗的患者给药如技术方案1到176任何一项中所述的肽、如技术方案202或205所述的免疫连接物、如技术方案206或207所述的药物组合物，或如技术方案178到182任何一项中所述的表达载体。Technical scheme 210: A method for suppressing a disease or symptom in a patient that is involved in CD38-expressing cells, the method comprising administering the peptide described in any one of technical schemes 1 to 176, such as technical scheme The immune linker described in 202 or 205, the pharmaceutical composition as described in technical scheme 206 or 207, or the expression vector as described in any one of technical schemes 178 to 182.

技术方案211：如技术方案209或210所述的方法，其中疾病或症状是风湿性关节炎。Technical solution 211: The method as described in technical solution 209 or 210, wherein the disease or symptom is rheumatoid arthritis.

技术方案212：如技术方案209或210所述的方法，其中疾病或症状是多发性骨髓瘤。Technical solution 212: The method as described in technical solution 209 or 210, wherein the disease or symptom is multiple myeloma.

技术方案213：如技术方案209到212任何一项中所述的方法，其中方法包括对患者给药一种或多种其它治疗性药剂。Technical solution 213: The method as described in any one of technical solutions 209 to 212, wherein the method includes administering one or more other therapeutic agents to the patient.

技术方案214：如技术方案213所述的方法，其中一种或多种其它治疗性药剂选自化疗药剂、抗炎症药剂、或免疫抑制和/或免疫调节药剂。Technical solution 214: The method as described in technical solution 213, wherein one or more other therapeutic agents are selected from chemotherapeutic agents, anti-inflammatory agents, or immunosuppressive and/or immunomodulatory agents.

技术方案215：如技术方案213所述的方法，其中一种或多种其它治疗性药剂选自施铂锭、吉非替尼、西妥昔单抗、利妥昔单抗、贝伐单抗、埃罗替尼、硼替佐米、沙利度胺、帕米膦酸盐、唑来膦酸、氯膦酸盐、利塞膦酸盐、伊班膦酸盐、依替膦酸盐、阿仑膦酸盐、替鲁膦酸盐、三氧化二砷、来那度胺、非格司亭、聚乙二醇化非格司亭、沙格司亭、辛二酰苯胺异羟肟酸和SCIO-469。Technical scheme 215: The method as described in technical scheme 213, wherein one or more other therapeutic agents are selected from platinum, gefitinib, cetuximab, rituximab, bevacizumab , erlotinib, bortezomib, thalidomide, pamidronate, zoledronic acid, clodronate, risedronate, ibandronate, etidronate, algae Lendronate, tiludronate, arsenic trioxide, lenalidomide, filgrastim, pegfilgrastim, sargragrastim, suberoylanilide hydroxamic acid, and SCIO-469.

技术方案216：一种在样品中检测CD38抗原或表达CD38的细胞存在的体外方法，包括：a)将样品与如技术方案1到176任何一项中 所述的肽在肽与CD38可形成复合物的条件下进行接触；及b)检测复合物的形成。Technical scheme 216: an in vitro method for detecting the presence of CD38 antigen or CD38-expressing cells in a sample, comprising: a) combining the sample with the peptide described in any one of technical schemes 1 to 176 where the peptide and CD38 can form a complex contacting under conditions of the substance; and b) detecting the formation of the complex.

技术方案217：如技术方案216所述的体外方法，其中所述肽是抗体。Technical scheme 217: The in vitro method as described in technical scheme 216, wherein the peptide is an antibody.

技术方案218：在含如技术方案1到176任何一项中所述的肽的样品中检测CD38抗原或表达CD38的细胞存在的试剂盒。Technical solution 218: A kit for detecting the presence of CD38 antigen or CD38-expressing cells in a sample containing the peptide described in any one of technical solutions 1 to 176.

技术方案219：检测患者CD38抗原或表达CD38的细胞存在的体内方法，包括：a)在肽与CD38可形成复合物的条件下给药如技术方案1到176任何一项中所述的肽；及b)检测形成的复合物。Technical solution 219: an in vivo method for detecting the presence of CD38 antigen or CD38-expressing cells in a patient, comprising: a) administering the peptide as described in any one of technical solutions 1 to 176 under the condition that the peptide and CD38 can form a complex; and b) detecting the complex formed.

技术方案220：如技术方案219所述的体内方法，其中所述肽是抗体。Technical scheme 220: the in vivo method as described in technical scheme 219, wherein the peptide is an antibody.

技术方案221：与如技术方案2、4或163到174任何一项中所述的肽结合的抗独特型抗体。Technical scheme 221: an anti-idiotypic antibody that binds to the peptide described in technical scheme 2, 4 or any one of 163 to 174.

技术方案222：如技术方案221所述的抗独特型抗体在检测样品中如技术方案2、4或163到174任何一项中所述的肽的水平方面的用途。Technical scheme 222: Use of the anti-idiotypic antibody as described in technical scheme 221 in detecting the level of the peptide as described in any one of technical schemes 2, 4 or 163 to 174 in a sample.

技术方案223：如技术方案221所述的抗独特型抗体在检测样品中抗CD38的人单克隆抗体水平方面的用途。Technical scheme 223: Use of the anti-idiotypic antibody as described in technical scheme 221 in detecting the level of human monoclonal antibody against CD38 in a sample.

术语“CD38”和“CD38抗原”此处可交换使用，而且包括任何由细胞天然表达的或在用CD38基因转染的细胞中表达的人CD38的变异体、同种型和种同源物。CD38的异名在本领域中包括ADP核糖基环化酶1、cADPr水解酶1、Cd38-rs1、环化ADP-核糖水解酶1、1-19、NIM-R5抗原。The terms "CD38" and "CD38 antigen" are used interchangeably herein and include any variants, isoforms and species homologues of human CD38 that are naturally expressed by cells or expressed in cells transfected with the CD38 gene. Synonyms for CD38 in the art include ADP ribosyl cyclase 1, cADPr hydrolase 1, Cd38-rsl, cyclizing ADP-ribohydrolase 1, 1-19, NIM-R5 antigen.

此处所述的术语肽是指CD38结合肽和非CD38肽，包括任何合适的肽，除非文中特别说明，可与术语多肽和蛋白通用；只要读者认为每种含各种氨基酸多聚体的分子具有显著差异，从而可形成本发明的单个技术方案即可(例如，含有多个多肽链的肽，例如抗体与单链抗体、肽免疫黏附剂或单链免疫肽显著不同)。因此，此处的术语肽可被理解为任何合适大小和组成(包括蛋白分子中的氨基酸数目和相互作用的了链的数目)的任何合适的肽。The term peptide described here refers to CD38 binding peptide and non-CD38 peptide, including any suitable peptide, unless otherwise specified in the text, it can be used generically with the term polypeptide and protein; as long as the reader thinks that each molecule containing various amino acid polymers Significant differences can form a single technical solution of the present invention (for example, peptides containing multiple polypeptide chains, such as antibodies and single-chain antibodies, peptide immunoadhesives or single-chain immune peptides are significantly different). Accordingly, the term peptide herein is to be understood as any suitable peptide of any suitable size and composition, including the number of amino acids in the protein molecule and the number of interacting chains.

另外，除非文中特别说明，此处所述的发明方法和组分中的肽可包括非天然存在和/或非L氨基酸残基。除非文中特别说明，术语肽进一步(如个别技术方案中的术语多肽和/或蛋白)也包括衍生的肽分子。简言 之，在本发明的文中，衍生物是在肽中的一个或多个氨基酸残基被化学修饰(例如烷基化、酰基化、酯化或胺化成)，或与一个或多个非氨基酸有机和/或无机原子或分子取代物(例如，聚乙烯二醇(PEG)基团、亲脂取代物(选择性地可与肽的氨基酸序列通过间隔区残基或基团，例如β-丙氨酸、γ-氨基丁酸(GABA)、UD-谷氨酸、琥珀酸等)、荧光、生物素、放射性核等)连接的肽，也可以或选择性地含有非必需、非天然存在和/或非L氨基酸残基，除非文中特别说明(但应再次意识到，这种衍生物其本身及其包括的分子可视为本发明的独立特征，将这类分子包括在肽的概念内是为了更方便地描述本发明，而不是表示在裸肽与这类衍生物之间存在任何等价关系)。这种氨基酸残基的非限定例子包括，例如2-氨基脂肪酸、3-氨基脂肪酸、β-丙氨酸、β-氨基丙酸、2-氨基丁酸、4-氨基丁酸、6-氨基己酸、2-氨基庚酸、2-氨基异丁酸、3-氨基异丁酸、2-氨基环己酸、2，4-二氨基丁酸、锁链素、2，2′-二氨基环己酸、2，3-二氨基丙酸、N-乙烷基甘氨酸、N-乙烷基天冬氨酸、羟基赖氨酸、异羟基赖氨酸、3-羟基脯氨酸、4-羟基脯氨酸、异锁链赖氨素、同分异构异亮氨酸、N-甲基甘氨酸、6-N-甲基赖氨酸、正缬氨酸、正亮氨酸、鸟氨酸和施德丁卤化的氨基酸。Additionally, unless otherwise indicated herein, the peptides in the inventive methods and compositions described herein may include non-naturally occurring and/or non-L amino acid residues. The term peptide further (such as the term polypeptide and/or protein in individual technical solutions) also includes derivatized peptide molecules, unless otherwise specified in the context. Briefly, in the context of the present invention, a derivative is a peptide in which one or more amino acid residues have been chemically modified (eg, alkylated, acylated, esterified or aminated), or combined with one or more non- Amino acid organic and/or inorganic atomic or molecular substituents (for example, polyethylene glycol (PEG) groups, lipophilic substituents (optionally with the amino acid sequence of the peptide via spacer residues or groups, such as β- Alanine, γ-aminobutyric acid (GABA, UD-glutamic acid, succinic acid, etc.), fluorescent, biotin, radionuclear, etc.) linked peptides that may also or optionally contain non-essential, non-naturally occurring and/or non-L amino acid residues, unless otherwise specified in the text (but again, it should be appreciated that such derivatives themselves and the molecules they comprise may be considered independent features of the invention, and such molecules are included within the concept of peptide It is for the purpose of describing the present invention more conveniently, and does not mean that there is any equivalence relationship between the naked peptide and such derivatives). Non-limiting examples of such amino acid residues include, for example, 2-amino fatty acid, 3-amino fatty acid, β-alanine, β-alanine, 2-aminobutyric acid, 4-aminobutyric acid, 6-aminohexanoic acid Acid, 2-aminoheptanoic acid, 2-aminoisobutyric acid, 3-aminoisobutyric acid, 2-aminocyclohexanoic acid, 2,4-diaminobutyric acid, desmosine, 2,2′-diaminocyclohexyl Acid, 2,3-Diaminopropionic acid, N-Ethylglycine, N-EthylAspartic Acid, Hydroxylysine, Isohydroxylysine, 3-Hydroxyproline, 4-Hydroxyproline Amino acid, iso-leucine, iso-isoleucine, N-methylglycine, 6-N-methyllysine, norvaline, norleucine, ornithine and styredine halogenated of amino acids.

抗原结合肽是指任何与部分给定抗原在细胞和/或生理条件下特异结合的肽，从而可在充分时间内诱导、促进、增强和/或调控与给抗原相关的生理功能；可通过此处所述的和/或本领域已知的ELISA、Western印迹或其它类似合适的蛋白结合技术和/或在相关时间内进行检测(例如至少约1 5分钟、至少约30分钟、至少约45分钟、至少约1小时、至少约2小时、至少约4小时、至少约6小时、至少约12小时、约1-24小时、约1-36小时、约1-48小时、约1-72小时、约1周或更长时间)。Antigen-binding peptide refers to any peptide that specifically binds to a given antigen under cellular and/or physiological conditions, thereby inducing, promoting, enhancing and/or regulating the physiological functions related to the given antigen in a sufficient time; ELISA, Western blot, or other similar suitable protein binding techniques described herein and/or known in the art and/or for detection within a relevant time period (e.g., at least about 15 minutes, at least about 30 minutes, at least about 45 minutes , at least about 1 hour, at least about 2 hours, at least about 4 hours, at least about 6 hours, at least about 12 hours, about 1-24 hours, about 1-36 hours, about 1-48 hours, about 1-72 hours, about 1 week or more).

CD38结合肽，或CD38BP，是与抗原CD38特异结合的抗原结合肽。在一个技术方案中，CD38BP与CD38的结合是通过使用实施例4所述的方法测定的。CD38-binding peptides, or CD38BPs, are antigen-binding peptides that specifically bind the antigen CD38. In one embodiment, the binding of CD38BP to CD38 is determined using the method described in Example 4.

术语免疫球蛋白是指一类结构相关的糖蛋白，由两对多肽链组成，一对为轻的(L)低分子量链，一对是重(H)链，所有四条链通过二硫键内部连接。免疫球蛋白的结构已被详细描述。参见，例如FundamentalImmunology，第七章(Paul，W.，ed.，2nd ed.Raven Press，N.Y.(1989))。简言之，每个重链典型地包括重链可变区(此处缩写为V_H)和重链恒 定区。重链恒定区典型地包括三个结构域，C_H1、C_H2和C_H3。每个轻链典型地包括轻链可变区(此处缩写为V_L)和轻链恒定区。轻链恒定区典型地包括一个结构域C_L。V_H和V_L区可进一步细分为高变区(或结构确定的环的序列和/或形式可高度变化的高变区)，也称为互补决定区(CDRs)，其中散布着更保守的称为框架区(FRs)的区域。The term immunoglobulin refers to a class of structurally related glycoproteins consisting of two pairs of polypeptide chains, one pair of light (L) low molecular weight chains and one pair of heavy (H) chains, all four chains are internally linked by disulfide bonds connect. The structure of immunoglobulins has been described in detail. See, eg, Fundamental Immunology, Chapter 7 (Paul, W., ed., 2nd ed. Raven Press, NY (1989)). Briefly, each heavy chain typically includes a heavy chain variable region (abbreviated herein as _VH ) and a heavy chain constant region. The heavy chain constant region typically includes three domains, _CH1 , _CH2 and _CH3 . Each light chain typically includes a light chain variable region (abbreviated herein as _VL ) and a light chain constant region. The light chain constant region typically includes one domain, _CL . The _VH and _VL regions can be further subdivided into hypervariable regions (or hypervariable regions in which the sequence and/or form of structurally defined loops can be highly variable), also called complementarity determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs).

每个V_H和V_L典型地包括3个CDRs和4个FRs，从氨基末端到羧基末端以如下顺序排列：FR1、CDR1、FR2、CDR2、FR3、CDR3、FR4(也参见Chothia and Lesk J.MoI.Biol.196，901-917(1987))。典型地，该区氨基酸残基的数目通过Kabat et al.，Sequences of ProteinsofImmunological Interest，第五版，Public Health Service，National InstitutesofHealth，Bethesda，MD.(1991)中所述的方法测定(诸如按Kabat中或根据此处Kabat进行编号的可变结构域残基这样的短语，是指用于重链可变结构域或轻链可变结构域的编号系统)。使用该编号系统，肽的实际线性氨基酸序列可含有更少的或额外的氨基酸，相当于可变结构域的FR或CDR的缩短或插入。例如，重链可变结构域可包括在V_H CDR2的残基52后有单个氨基酸的插入(根据Kabat的残基52a)和在重链FR残基82后有插入的残基(例如根据Kabat的残基82a、82b和82c等)。对给定的抗体，残基的Kabat编号可通过将抗体序列的同源区与“标准的”Kabat编号序列比对后而确定。Each _VH and _VL typically includes 3 CDRs and 4 FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 (see also Chothia and Lesk J. MoI. Biol. 196, 901-917 (1987)). Typically, the number of amino acid residues in this region is determined by the method described in Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, Public Health Service, National Institutes of Health, Bethesda, MD. (1991) (such as in Kabat Or phrases such as variable domain residues numbered according to Kabat here refer to the numbering system used for heavy chain variable domains or light chain variable domains). Using this numbering system, the actual linear amino acid sequence of a peptide may contain fewer or additional amino acids corresponding to shortenings or insertions of FRs or CDRs of the variable domains. For example, the heavy chain variable domain may include a single amino acid insertion after residue 52 of the _VH CDR2 (residue 52a according to Kabat) and an insertion after residue 82 of the heavy chain FR (e.g., according to Kabat Residues 82a, 82b and 82c of , etc.). For a given antibody, the Kabat numbering of residues can be determined by aligning the homologous regions of the antibody sequence with "standard" Kabat numbering sequences.

本发明文中的抗体(Ab)是指免疫球蛋白分子，免疫球蛋白分子的一部分或其衍生物，具有在典型地生理条件下可与抗原在足够的时间内，例如至少约30分钟，至少约45分钟，至少约1小时，至少约2小时，至少约4小时，至少约8小时，至少约12小时，至少约24小时或以上，至少约48小时或以上，至少约3、4、5、6、7或更多天等，或任何其它相关功能性确定的时期(例如足够诱导、促进、增强和/或调节与抗体结合抗原相关的生理应答)内与抗原特异结合的能力。Antibody (Ab) in the context of the present invention refers to an immunoglobulin molecule, a part of an immunoglobulin molecule or a derivative thereof, which has the ability to interact with an antigen in a sufficient time under typical physiological conditions, such as at least about 30 minutes, at least about 45 minutes, at least about 1 hour, at least about 2 hours, at least about 4 hours, at least about 8 hours, at least about 12 hours, at least about 24 hours or more, at least about 48 hours or more, at least about 3, 4, 5, The ability to specifically bind to an antigen within 6, 7 or more days, etc., or any other relevant functionally defined period of time (eg, sufficient to induce, promote, enhance and/or modulate a physiological response associated with antibody binding to the antigen).

免疫球蛋白分子的重链和轻链可变区含有与抗原相互作用的结合结构域。抗体(Abs)的恒定区可介导免疫球蛋白与宿主组织或因子，包括免疫系统的各种细胞(例如效应细胞)和典型的补体系统的第一成分(Clq)的结合。The variable regions of the heavy and light chains of immunoglobulin molecules contain binding domains that interact with antigens. The constant regions of antibodies (Abs) mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (eg, effector cells) and typically the first component (Clq) of the complement system.

抗CD38抗体可以为双特异性抗体、双功能抗体或相似分子(参见，例如PNAS USA90(14)，6444-8(1993)描述的双功能抗体)。实际 上，本发明提供的双特异性抗体，双功能抗体等可与除CD38部分外的任何其它合适的靶结合。Anti-CD38 antibodies may be bispecific antibodies, bifunctional antibodies or similar molecules (see, eg, bifunctional antibodies described in PNAS USA 90(14), 6444-8 (1993)). In fact, the bispecific antibodies, diabodies, etc. provided by the present invention can bind to any other suitable target except the CD38 portion.

如上所述，除非文中特别说明或明确矛盾，此处术语抗体包括具有特异结合抗原能力的抗体部分。已表明抗体的抗原结合功能可通过全长抗体的片段来完成。结合片段的例子包括在术语“抗体”中，包括(i)Fab片段，含有V_L、V_H、C_L和C_H1结构域的单价片段；(ii)F(ab)₂ 和F(ab′)₂片段，含2个Fab在铰链区通过二硫键连接的片段的双价片段；(iii)主要由V_H和C_H1结构域组成的Fd片段；(iv)主要由抗体的单个臂的V_L和V_H结构域组成的Fv片段；(v)dAb片段(Ward et al.，Nature 341，544-546(1989))，它主要由V_H结构域组成；(vi)独立的互补决定区(CDR)1，及(vii)两个或单个独立的CDRs的组合，可选择性地通过合成的连接物连接。另外，虽然Fv片段、V_L和V_H的两个结构域是由不同的基因编码的，但他们可通过重组方法，用可使其V_L 和V_H区形成单价分子而作为单个蛋白链的合成的连接物连接(已知作为单链抗体或单链Fv(scFv)，参见例如，Bird et al.，Science 242，423-426(1988)and Hustonet al.，PNAS USA 85，5879-5883(1988))。这种单链抗体在术语抗体的范围内，除非文中特别说明或明确提示。其它类型的单链抗体，例如双功能抗体包括在术语抗体的范围内。虽然这种片段通常包括在抗体的含义中，但它们所有每个都是本发明的独特特征，表现不同的生物学特性和用途。这些和其它有用的抗体片段在本文进一步讨论。As noted above, unless otherwise stated or clearly contradicted by context, the term antibody herein includes that portion of an antibody that has the ability to specifically bind an antigen. It has been shown that the antigen-binding function of antibodies can be performed by fragments of full-length antibodies. Examples of binding fragments included within the term "antibody" include (i) Fab fragments, monovalent fragments containing _VL , _VH , _CL and _CH1 domains; (ii) F(ab) ₂ and F(ab ') ₂ fragments, a bivalent fragment containing 2 Fab fragments connected by disulfide bonds at the hinge region; (iii) Fd fragment mainly composed of _VH and _CH1 domains; (iv) mainly composed of a single antibody fragment Fv fragment composed of the V _L and V _H domains of the arm; (v) dAb fragment (Ward et al., Nature 341, 544-546 (1989)), which is mainly composed of the V _H domain; (vi) independent The complementarity determining region (CDR) 1, and (vii) a combination of two or single individual CDRs, optionally joined by a synthetic linker. In addition, although the two structural domains of the Fv fragment, V _L and V _H , are encoded by different genes, they can be used as a single protein chain by recombination methods that allow their V _L and V _H regions to form monovalent molecules. Synthetic linker ligation (known as single-chain antibody or single-chain Fv (scFv), see, for example, Bird et al., Science 242, 423-426 (1988) and Hustone et al., PNAS USA 85, 5879-5883 ( 1988)). Such single chain antibodies are within the scope of the term antibody unless otherwise specified or explicitly suggested by the context. Other types of single chain antibodies, such as diabodies are included within the scope of the term antibody. While such fragments are generally included within the meaning of antibodies, each of them is a unique feature of the invention, exhibiting different biological properties and uses. These and other useful antibody fragments are discussed further herein.

也应理解术语抗体也通常包括多克隆抗体、单克隆抗体(mAbs)，抗体类似的多肽，例如通过任何已知技术，例如酶切割、肽合成和重组技术制备的嵌合抗体和人源化抗体，抗体的抗独特型(抗Id)抗体和具有与抗原特异结合的能力的抗体片段(抗原结合片段)。产生的抗体具有任何同种型。It is also understood that the term antibody also generally includes polyclonal antibodies, monoclonal antibodies (mAbs), antibody-like polypeptides, such as chimeric and humanized antibodies prepared by any known technique, such as enzymatic cleavage, peptide synthesis and recombinant techniques , anti-idiotypic (anti-Id) antibodies of antibodies and antibody fragments (antigen-binding fragments) having the ability to specifically bind to an antigen. Antibodies produced can be of any isotype.

抗CD38抗体是如上所述的抗体，它可与抗原CD38特异结合。An anti-CD38 antibody is an antibody as described above that specifically binds to the antigen CD38.

术语“抗原决定簇”是指可与抗体特异结合的蛋白决定区。抗原决定簇通常由分子的有化学活性的表面基团，例如氨基酸或糖侧链组成，通常具有特定的3个二维结构特征，及特定的电荷特征。可区分构象和非构象抗原决定簇，因为在存在变性溶剂时，前者的结合消失，而后者的结合不消失。抗原决定簇可含有直接参与结合的氨基酸残基(也称为 抗原决定簇的免疫优势组分)和其它氨基酸残基，它们不直接参与结合，例如通过特异的抗原结合肽而被有效封闭的氨基酸残基(即，氨基酸残基在特异的抗原结合肽的覆盖区)。The term "antigenic determinant" refers to a determinant region of a protein that can specifically bind to an antibody. Antigenic determinants usually consist of chemically active surface groups of molecules, such as amino acids or sugar side chains, and usually have specific three-dimensional structural characteristics and specific charge characteristics. A distinction can be made between conformational and non-conformational epitopes, since binding of the former but not the latter disappears in the presence of denaturing solvents. An antigenic determinant may contain amino acid residues that are directly involved in binding (also known as the immunodominant component of the epitope) and other amino acid residues that are not directly involved in binding, such as amino acids effectively blocked by specific antigen-binding peptides residues (ie, amino acid residues in the footprint of a specific antigen-binding peptide).

术语“双特异性分子”包括任何试剂，例如蛋白、肽和蛋白或肽的复合物，它具有两个不同的结合特异性。例如，该分子可与(a)细胞表面的抗原和(b)效应细胞编码的Fc受体结合或相互作用。术语“多特异性分子”包括任何试剂，例如蛋白、肽和蛋白或肽的复合物，它具有多于两个不同的结合特异性。例如，该分子可与(a)细胞表面的抗原，(b)效应细胞编码的Fc受体和(c)至少一种其它组分结合或相互作用。因此，本发明包括，但不限定于，针对CD3 8和其它细胞表面抗原或靶，例如效应细胞的Fc受体的双特异性、三特异性、四特异性和其它多特异性分子。The term "bispecific molecule" includes any agent, such as a protein, a peptide, and a complex of proteins or peptides, which has two different binding specificities. For example, the molecule can bind or interact with (a) an antigen on the cell surface and (b) an Fc receptor encoded by an effector cell. The term "multispecific molecule" includes any agent, such as proteins, peptides and complexes of proteins or peptides, which has more than two different binding specificities. For example, the molecule can bind or interact with (a) an antigen on the cell surface, (b) an Fc receptor encoded by an effector cell, and (c) at least one other component. Thus, the invention includes, but is not limited to, bispecific, trispecific, tetraspecific and other multispecific molecules directed against CD38 and other cell surface antigens or targets, such as Fc receptors of effector cells.

术语“双特异性抗体”包括任何双特异性分子的抗CD38抗体。术语“双特异性抗体”也包括双功能抗体。双功能抗体是二价的双特异性抗体，其中V_H和V_L结构域在单个多肽链上表达，但通过同一链上的两个结构域间的较短的不能使其配对的连接物进行连接，从而使结构域与另一条链的互补结构域配对，从而得到两个抗原结合位点(参见，例如Holliger，P.et al.，PNAS USA 90，6444-6448(1993)、Poljak，RJ.et al.，Structure 2，1121-1123(1994))。The term "bispecific antibody" includes anti-CD38 antibodies of any bispecific molecule. The term "bispecific antibody" also includes bifunctional antibodies. Diabodies are bivalent, bispecific antibodies in which the _VH and _VL domains are expressed on a single polypeptide chain, but through a shorter linker between the two domains on the same chain that does not enable their pairing. Linking such that the domains are paired with complementary domains of another chain, resulting in two antigen-binding sites (see, e.g., Holliger, P. et al., PNAS USA 90, 6444-6448 (1993), Poljak, RJ et al., Structure 2, 1121-1123 (1994)).

此处所用术语“效应细胞”是指参与免疫应答的效应阶段的免疫细胞，与免疫应答的认知和激活阶段相对应。示例的免疫细胞包括骨髓或淋巴来源的细胞，例如，淋巴细胞(例如B细胞和T细胞，包括溶细胞的T细胞(CTLs))，杀伤细胞、自然杀伤细胞、巨噬细胞、单核细胞、嗜曙红细胞、嗜中性粒细胞、分叶核白细胞、粒细胞、肥大细胞和嗜碱细胞。某些效应细胞表达特异的Fc受体，并具有特定的免疫功能。在某些技术方案中，效应细胞可诱导抗体依赖的细胞毒素作用(ADCC)，例如嗜中性粒细胞可诱导ADCC。例如，表达FcR的单核细胞、巨噬细胞参与特定的靶细胞杀伤，并向免疫系统的其它组分呈递抗原，或与呈递抗原的细胞结合。在某些技术方案中，效应细胞可吞噬靶抗原、靶细胞或微生物。效应细胞上特定的FcR的表达可通过体液因子，例如细胞因子来调控。例如，已发现FcγRI的表达可被干扰素γ(IFN-γ)和/或G-CSF上调。这种增强的表达可提高含FcγRI的细胞针对靶的细胞毒活性。效应细胞可吞噬或裂解靶抗原或靶细胞。The term "effector cells" as used herein refers to immune cells that participate in the effector phase of an immune response, corresponding to the cognitive and activation phases of the immune response. Exemplary immune cells include cells of myeloid or lymphoid origin, e.g., lymphocytes (e.g., B cells and T cells, including cytolytic T cells (CTLs)), killer cells, natural killer cells, macrophages, monocytes, Eosinophils, neutrophils, segmented leukocytes, granulocytes, mast cells, and basophils. Certain effector cells express specific Fc receptors and have specific immune functions. In certain embodiments, the effector cells induce antibody-dependent cytotoxicity (ADCC), eg, neutrophils induce ADCC. For example, FcR-expressing monocytes, macrophages participate in specific target cell killing, and present antigens to other components of the immune system, or bind to cells presenting antigens. In some embodiments, the effector cells can phagocytose target antigens, target cells or microorganisms. The expression of specific FcRs on effector cells can be regulated by humoral factors, such as cytokines. For example, it has been found that the expression of FcγRI can be upregulated by interferon gamma (IFN-γ) and/or G-CSF. This enhanced expression increases the cytotoxic activity of FcyRI-containing cells against targets. Effector cells can phagocytose or lyse target antigens or target cells.

此处所用术语“人抗体”包括具有来源于人胚系免疫球蛋白序列的可变区和恒定区的抗体。本发明的人抗体也包括不被人胚系免疫球蛋白序列编码的氨基酸残基(例如，通过体外随机或定点突变或体内细胞突变引入的突变)。但此处所用术语“人抗体”不包括其来源于其它哺乳动物种，例如鼠的CDR序列移植到人的阅读框序列中的抗体。此处所用人抗体是“来源于”特定的胚系序列，如果抗体是从使用人免疫球蛋白序列的系统，例如通过免疫携带人免疫球蛋白基因的转基因鼠，或通过筛选人免疫球蛋白基因库而获得的，则筛选的人抗体的氨基酸序列与由胚系V_H或V_L可变区基因片段编码的氨基酸序列的同一性至少为90％，例如至少为95％，例如至少为96％，例如至少为97％，例如至少为98％，或例如至少为99％。典型地，来源于特定的人胚系V_H或V_L 可变区基因片段序列的人抗体与由胚系免疫球蛋白基因编码的氨基酸序列相比较，具有不超过10个氨基酸的差异，例如不超过5，例如不超过4、3、2或1个氨基酸差异。The term "human antibody" as used herein includes antibodies having variable and constant regions derived from human germline immunoglobulin sequences. The human antibodies of the invention also include amino acid residues not encoded by human germline immunoglobulin sequences (eg, mutations introduced by random or site-directed mutagenesis in vitro or cellular mutation in vivo). However, the term "human antibody" as used herein does not include antibodies whose CDR sequences derived from other mammalian species, such as mice, are grafted into human reading frame sequences. A human antibody as used herein is "derived" from a particular germline sequence if the antibody is obtained from a system using human immunoglobulin sequences, for example by immunization of transgenic mice carrying human immunoglobulin genes, or by screening human immunoglobulin gene repertoires If obtained, the amino acid sequence of the screened human antibody is at least 90%, such as at least 95%, such as at least 96%, identical to the amino acid sequence encoded by the germline _VH or _VL variable region gene segment, For example at least 97%, such as at least 98%, or such as at least 99%. Typically, a human antibody derived from a specific human germline _VH or _VL variable region gene sequence differs by no more than 10 amino acids from the amino acid sequence encoded by a germline immunoglobulin gene, e.g., no More than 5, such as no more than 4, 3, 2 or 1 amino acid difference.

嵌合抗体是含有一个或多个来源于一个抗体的区域和一个或多个来源于一个或多个来自另一个物种的其它抗体的区域的抗体。单价嵌合抗体是将嵌合的H链通过二硫键与嵌合的L链连接而形成的二聚体(HL))。二价嵌合抗体是两个HL二聚体通过至少一个二硫键连接而形成的四聚体(H₂L₂)。也可产生多价嵌合抗体，例如，通过使CH区寡聚化(例如来源于IgM H链或μ链)。典型地，嵌合抗体是指其重链和/或轻链部分与来源于特定物种的抗体的对应序列同一或同源的或属于特定的抗体类型或亚类，而链的其余部分与另一物种的抗体的对应序列同一或同源或属于另一种抗体类型或亚类的抗体及这种抗体的片段，只要它们具有所需的生物活性即可(参见，例如US 4,816,567和Morrison et al.，PNAS USA 81.，6851-6855(1984))。嵌合抗体通过本领域已知的重组过程产生(参见，例如Cabilly etal.，PNAS USA 81，3273-3277(1984)、Morrison et al.，PNAS USA 8JL 6851-6855(1984)、Boulianne et al.，Nature 312，643-646(1984)、EP 125023，Neuberger etal.，Nature314，268-270(1985)、EP 171496，EP173494、WO86/01533，EP 184187，Sahagan et al.，J.Immunol.137.1066-1074(1986)、WO87/02671，Liu et al.，PNAS USA 84，3439-3443(1987)、Sun et al.， PNAS USA 84，214-218(1987)、Better et al.，Science 240，1041-1043(1988)and Harlow et al.，Antibodies：A Laboratory Manual，Cold SpringHarborLaboratory Press，Cold Spring Harbor，N.Y.，(1988))。Chimeric antibodies are antibodies that contain one or more regions derived from one antibody and one or more regions derived from one or more other antibodies from another species. A monovalent chimeric antibody is a dimer (HL) formed by linking a chimeric H chain and a chimeric L chain through a disulfide bond. Bivalent chimeric antibodies are tetramers ( _H2L2 ) of _two HL dimers linked by at least one disulfide bond. Multivalent chimeric antibodies can also be produced, for example, by oligomerization of the CH region (eg, derived from IgM H chain or μ chain). Typically, a chimeric antibody refers to a heavy chain and/or light chain portion of which is identical or homologous to the corresponding sequence of an antibody derived from a specific species or belongs to a specific antibody type or subclass, while the rest of the chain is identical to another Species identical or homologous to the corresponding sequence of an antibody or belonging to another antibody type or subclass, and fragments of such antibodies, so long as they possess the desired biological activity (see, for example, US 4,816,567 and Morrison et al. , PNAS USA 81., 6851-6855 (1984)). Chimeric antibodies are produced by recombinant procedures known in the art (see, e.g., Cabilly et al., PNAS USA 81, 3273-3277 (1984), Morrison et al., PNAS USA 8JL 6851-6855 (1984), Boulianne et al. , Nature 312,643-646 (1984), EP 125023, Neuberger et al., Nature 314, 268-270 (1985), EP 171496, EP173494, WO86/01533, EP 184187, Sahagan et al., J.Immunol.137.1066- 1074(1986), WO87/02671, Liu et al., PNAS USA 84, 3439-3443(1987), Sun et al., PNAS USA 84, 214-218(1987), Better et al., Science 240, 1041 -1043 (1988) and Harlow et al., Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, (1988)).

人源化抗体是来源于非人物种的抗体，其中将阅读框和重链及轻链恒定区的特定氨基酸突变，从而可避免或消除在人中的免疫应答。非人(例如鼠)抗体的人源化形式是含有来源于非人免疫球蛋白最小序列的嵌合抗体。对大部分而言，人源化抗体是其受体的高变区残基被具有所需抗原结合特性，例如特异性和亲和性的非人物种(供体抗体)，例如小鼠、大鼠、兔或非人灵长类的高变区残基替代的人免疫球蛋白(受体抗体)。在某些情况下，人免疫球蛋白的Fv阅读框区域(FR)残基被相应的非人残基替代。另外，人源化抗体可含有受体抗体或供体抗体中没有的残基。这些修饰可进一步优化抗体的功能。通常，人源化抗体实质含有至少一个，典型地两个可变结构域，其中所有或实质所有高变区的环对应非人免疫球蛋白，或所有或实质所有FR区域是人免疫球蛋白的序列。人源化抗体选择性地也含有至少一部分免疫球蛋白的恒定区(Fc)，典型地是人免疫球蛋白的恒定区。详细内容参见Jones et al.，Nature 321，522-525(1986)、Riechmann et al.，Nature 332，323-329(1988)and Presta，Curr.Op.Struct.Biol.2，593-596(1992)。Humanized antibodies are antibodies derived from a non-human species in which specific amino acids in the reading frame and constant regions of the heavy and light chains have been mutated so that an immune response in humans can be avoided or eliminated. Humanized forms of non-human (eg, murine) antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin. For the most part, a humanized antibody is a non-human species (donor antibody) whose recipient hypervariable region residues have been modified to possess the desired antigen-binding properties, such as specificity and affinity (donor antibody), such as mouse, rat Human immunoglobulin (recipient antibody) substituted with hypervariable region residues from , rabbit or non-human primate. In certain instances, Fv reading frame region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. In addition, humanized antibodies can contain residues that are not found in either the recipient antibody or the donor antibody. These modifications can further optimize the function of the antibody. In general, humanized antibodies contain substantially at least one, typically two, variable domains in which all or substantially all of the loops of the hypervariable regions correspond to non-human immunoglobulins, or in which all or substantially all FR regions are of human immunoglobulins sequence. A humanized antibody optionally also will contain at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For details, see Jones et al., Nature 321, 522-525 (1986), Riechmann et al., Nature 332, 323-329 (1988) and Presta, Curr.Op.Struct.Biol.2, 593-596 (1992 ).

此处所用“单克隆抗体”或“单克隆抗体组分”是指单分子组分的抗体分子的制备。单克隆抗体组分具有单一的特定抗原决定簇的结合特异性和亲和性。因此，术语“人单克隆抗体”是指具有来源于人胚系免疫球蛋白序列的可变区和恒定区，并具有单一的结合特异性的抗体。人单克隆抗体可通过杂交瘤产生，包括从具有包含人重链转基因和轻链转基因的基因组的转基因或转染色体非人动物，例如转基因鼠获得B细胞与永生化细胞融合。单克隆抗体可缩写为mAb。As used herein, "monoclonal antibody" or "monoclonal antibody composition" refers to the preparation of antibody molecules of single molecular composition. Monoclonal antibody components have a single binding specificity and affinity for a particular antigenic determinant. Thus, the term "human monoclonal antibody" refers to an antibody having variable and constant regions derived from human germline immunoglobulin sequences and having a single binding specificity. Human monoclonal antibodies can be produced by hybridomas, including the fusion of B cells obtained from a transgenic or transchromosomal non-human animal, such as a transgenic mouse, with a genome comprising a human heavy chain transgene and a light chain transgene, with immortalized cells. A monoclonal antibody may be abbreviated as mAb.

此处所用术语“重组人抗体”包括所有通过重组方法制备、表达、产生或分离的人抗体，例如(a)从含有人免疫球蛋白基因的转基因或转染色体动物(例如鼠)中分离的抗体或制备的杂交瘤(在本文其它部分描述)，(b)从转化的以表达抗体的宿主细胞，例如转染瘤分离的抗体，(c)从重组的、组合人抗体库分离的抗体，及(d)通过任何其它涉及将人免疫球蛋白基因序列与其它DNA序列拼接的方法制备、表达、产生或分离的抗体。这种重组的人抗体具有来源于人胚系免疫球蛋 白序列的可变区和恒定区。但在某些技术方案中，这种重组人抗体可进行体外突变(或当对人Ig序列使用动物转基因时，为体内体细胞突变)，因此，重组抗体的V_H和V_L区的氨基酸序列，当它们来源于人胚系V_H 和V_L序列并与之相关时，可以是体内人抗体胚系库天然不存在的序列。The term "recombinant human antibody" as used herein includes all human antibodies prepared, expressed, produced or isolated by recombinant methods, for example (a) antibodies isolated from transgenic or transchromosomal animals (such as murines) containing human immunoglobulin genes or prepared hybridomas (described elsewhere herein), (b) antibodies isolated from host cells transformed to express antibodies, such as transfectomas, (c) antibodies isolated from recombinant, combinatorial human antibody repertoires, and (d) Antibodies prepared, expressed, produced or isolated by any other method involving the splicing of human immunoglobulin gene sequences with other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in some technical schemes, this recombinant human antibody can undergo in vitro mutation (or in vivo somatic mutation when animal transgenes are used for human Ig sequences), therefore, the amino acid sequences of the _VH and _VL regions of the recombinant antibody , when derived from and related to human germline _VH and _VL sequences, may be sequences that do not naturally occur in the human antibody germline repertoire in vivo.

此处所用“异源抗体”定义为涉及转基因非人生物产生的抗体。该术语是指与在不由非人动物所组成的生物中，通常是在来源于除了转基因非人动物外的某个物种中所发现的抗体具有相对应的氨基酸序列的抗体。"Heterologous antibody" as used herein is defined to refer to antibodies produced by a genetically modified non-human organism. The term refers to an antibody that has an amino acid sequence corresponding to an antibody found in an organism that does not consist of a non-human animal, usually a species derived from a species other than a transgenic non-human animal.

此处所用“分离的抗体”是指实质不含有其它具有不同的抗原特异性的抗体(例如，与CD38特异结合的分离抗体实质不含有与除CD38外的抗原特异结合的抗体)的抗体。但与人CD38的抗原决定簇、同种型或变异体特异结合的分离抗体可与其它相关的抗原，例如来源于其它物种(例如CD38物种同源体)具有交叉反应性。另外，分离抗体实质不含有其它细胞物质和/和化学制品。在本发明的一个技术方案中，具有不同特异性的“分离的”单克隆抗体的组合可含有明确的成分。As used herein, "isolated antibody" refers to an antibody that is substantially free of other antibodies with different antigen specificities (eg, an isolated antibody that specifically binds to CD38 does not contain substantially antibodies that specifically bind to antigens other than CD38). However, an isolated antibody that specifically binds to an epitope, isoform or variant of human CD38 may have cross-reactivity with other related antigens, eg, from other species (eg CD38 species homologues). Additionally, an isolated antibody is substantially free of other cellular material and/and chemicals. In one embodiment of the invention, the combination of "isolated" monoclonal antibodies with different specificities may contain distinct components.

此处所用“特异结合”是指诸如抗体这样的抗原结合肽，结合到预定的抗原上。典型地，当通过表面等离子共振(SPR)技术在BIAcore 3000仪器上，以重组CD38为配体，抗体为分析物进行测定时，诸如抗体这样的抗原结合肽的结合亲和性相当于K_D为10^-7M或更少，例如约10^-8M或更少，例如约10^-9M或更少，约10^-10M或更少，约10^-11M或更少。抗原结合肽可与预定抗原以相当于的K_D，比其与除预定蛋白或密切相关的抗原之外的非特异抗原(例如BSA、酪蛋白)的结合亲和性低至少10倍，例如至少低100倍，例如至少低1000倍，例如至少低10,000倍，例如至少低1000,000倍的亲和力结合。具有更低的亲和力的量依赖于抗原结合肽的K_D，因此当抗原结合肽的K_D很低时(即抗原结合肽是高度特异的)，则与抗原亲和力的量比与非特异抗原亲和力至少低10,000倍。此处所用短语“识别抗原的抗原结合肽”和“抗原特异的抗原结合肽”可与术语“与抗原特异结合的抗原结合肽”互换使用。同样，此处所用短语“识别抗原的抗体”和“抗原特异的抗体”可与术语“与抗原特异结合的抗体”互换使用。As used herein, "specifically binds" means that an antigen-binding peptide, such as an antibody, binds to a predetermined antigen. Typically, an antigen-binding peptide such as an antibody has a binding affinity equivalent to a _K of 10 ⁻⁷ M or less, such as about 10 ⁻⁸ M or less, such as about 10 ⁻⁹ M or less, about 10 ⁻¹⁰ M or less, about 10 ⁻¹¹ M or less. The antigen-binding peptide may bind to the intended antigen with an equivalent _KD that is at least 10-fold lower than its binding affinity to non-specific antigens (e.g., BSA, casein) other than the intended protein or closely related antigens, e.g., at least 100-fold lower, such as at least 1000-fold lower, such as at least 10,000-fold lower, such as at least 1000,000-fold lower affinity binding. Quantities with lower affinities are dependent on the KD of the antigen-binding peptide, so when the _KD of the antigen-binding _peptide is low (i.e., the antigen-binding peptide is highly specific), then the quantity with affinity for the antigen is more specific than the quantity with affinity for the non-specific antigen At least 10,000 times lower. As used herein, the phrases "antigen-binding peptide that recognizes an antigen" and "antigen-specific antigen-binding peptide" are used interchangeably with the term "antigen-binding peptide that specifically binds to an antigen." Also, as used herein, the phrases "antibody that recognizes an antigen" and "antigen-specific antibody" are used interchangeably with the term "antibody that specifically binds an antigen."

此处所用术语“k_d”(sec^-1)是指特定的抗体抗原相互作用的解离平衡速率常数。所述值也指k_off值。The term "k _d " (sec ^-1 ) as used herein refers to the dissociation equilibrium rate constant for a particular antibody-antigen interaction. Said values are also referred to as k _off values.

此处所用术语“k_a”(M^-1×sec^-1)是指特定的抗体抗原相互作用的结合平衡速率常数。The term " _ka " (M ^-1 xsec ^-1 ) as used herein refers to the equilibrium rate constant for binding of a particular antibody-antigen interaction.

此处所用术语“k_D”(M)是指特定的抗体抗原相互作用的解离平衡常数。The term " _kD " (M) as used herein refers to the dissociation equilibrium constant for a particular antibody-antigen interaction.

此处所用术语“K_A”(M^-1)是指特定的抗体抗原相互作用的结合平衡常数，通过k_a除以k_d而得到。The term "KA" ( _M ^-1 ) as used herein refers to the binding equilibrium constant for a particular antibody-antigen interaction, obtained by dividing _ka by _kd .

此处所用“同种型”是指由重链恒定区基因编码的抗体类型(例如IgG1、IgG2、IgG3、IgG4、IgD、IgA、IgE、或IgM)。"Isotype" as used herein refers to the antibody class (eg, IgGl, IgG2, IgG3, IgG4, IgD, IgA, IgE, or IgM) encoded by the heavy chain constant region genes.

此处所用“同种型转换”是指抗体的类型或同种型从一种免疫球蛋白类型转变为一种其它的免疫球蛋白类型的现象。"Isotype switching" as used herein refers to the phenomenon in which the class or isotype of an antibody changes from one immunoglobulin class to another.

此处所用“非转换的同种型”是指在无同种型转换发生时产生的重链的同种型类型；编码非转换同种型的CH基因典型地是紧接功能性重组VDJ基因下游的第一个CH基因。同种型转换分为典型的或非典型的同种型转换。典型的同种型转换通过至少转基因的一个转换序列区参与的重组事件发生。非典型的同种型转换可发生在例如人σμ和人∑μ(δ-相关的缺失)间的同源重组。选择性的非典型转换机制，例如转基因内和/或染色体内重组可发生，并实现同种型转换。"Non-switched isoform" as used herein refers to the isotype type of the heavy chain produced when no isotype switching occurs; the CH gene encoding the non-switched isoform is typically followed by a functionally recombinant VDJ gene The first CH gene downstream. Isotype switching is classified as typical or atypical isotype switching. Typically isotype switching occurs through recombination events involving at least one switch sequence region of the transgene. Atypical isotype switching can occur, for example, by homologous recombination between human σμ and human Σμ (δ-associated deletion). Alternative atypical switching mechanisms such as intra-transgenic and/or intra-chromosomal recombination can occur and achieve isotype switching.

此处所用术语“转换序列”是指那些负责转换重组的DNA序列。“转换供体”序列典型地是μ转换区，是在转换重组过程中被删除的恒定区的5′(即上游)。“转换受体”区是被删除的恒定区和替代的恒定区(例如γ、ε等)之间。由于在发生重组的地方没有特定的位点，因此典型地，最终的基因序列不能从构建体来预测。The term "switch sequence" as used herein refers to those DNA sequences responsible for switch recombination. The "switch donor" sequence is typically the mu switch region, 5' (ie, upstream) of the constant region that is deleted during switch recombination. A "switch acceptor" region is between the deleted constant region and the replacement constant region (eg gamma, epsilon, etc.). Since there is no specific site where recombination occurs, typically the final gene sequence cannot be predicted from the construct.

此处所用“糖基化模式”定义为与蛋白，更具体地与免疫球蛋白(抗体)蛋白共价连接的碳水化合物单元的模式。异源抗体的糖基化模式的特征在于与由非人转基因动物物种产生的抗体上天然存在的糖基化模式实质上是相似的，本领域普通人员会意识到异源抗体的糖基化模式与所述非人转基因动物物种的糖基化模式比与转基因的CH基因来源的物种具有更大的相似性。As used herein, "glycosylation pattern" is defined as the pattern of carbohydrate units covalently attached to proteins, more specifically to immunoglobulin (antibody) proteins. The glycosylation pattern of the heterologous antibody is characterized by being substantially similar to the glycosylation pattern naturally occurring on the antibody produced by the non-human transgenic animal species, and those of ordinary skill in the art will recognize the glycosylation pattern of the heterologous antibody The glycosylation pattern is more similar to the species of the non-human transgenic animal than to the species from which the transgenic CH gene was derived.

此处所用术语“天然存在的”用于某受试体时是指该受试体是在自然界发现的。例如，可从自然界来源分离的，没有在实验室人工改变的生物(包括病毒)中存在的多肽或多聚核酸序列是天然存在的。As used herein, the term "naturally occurring" when applied to a subject means that the subject is found in nature. For example, a polypeptide or polynucleic acid sequence that is isolated from a natural source and that is not found in laboratory-modified organisms, including viruses, is naturally occurring.

此处所用术语“重排的”是指重链或轻链免疫球蛋白座位的构象， 其中在分别编码完整的V_H或V_L结构域的构象中，V片段位于紧邻D-J或J片段。重组的免疫球蛋白(抗体)基因座位可通过与胚系DNA比较来鉴定；重组的座位具有至少一个七聚物/九聚物同源元件。The term "rearranged" as used herein refers to a conformation of a heavy or light chain immunoglobulin locus wherein the V segment is located immediately adjacent to the DJ or J segment in a conformation encoding a complete _VH or _VL domain, respectively. Recombined immunoglobulin (antibody) gene loci can be identified by comparison to germline DNA; recombinant loci have at least one heptamer/nonamer homologous element.

此处所用术语“未重排的”或“胚系构象”参照V片段而言，是指其中V片段不重组以与D或J片段紧邻的构象。The term "unrearranged" or "germline conformation" as used herein with reference to the V segment refers to a conformation in which the V segment does not recombine to be in close proximity to the D or J segment.

此处所用术语“核酸分子”是指DNA分子和RNA分子。核酸分子可以是单链的或双链的，但优选地是双链DNA。核酸可存在于全细胞、细胞裂解物中或是部分纯化或基本纯化的形式。The term "nucleic acid molecule" as used herein refers to DNA molecules and RNA molecules. Nucleic acid molecules can be single-stranded or double-stranded, but are preferably double-stranded DNA. Nucleic acids may be present in whole cells, cell lysates, or in partially or substantially purified form.

当通过标准技术，包括碱/SDS处理、CsCl成带、柱色谱、琼脂糖凝胶电泳和其它本领域公知的技术从其它细胞成分或诸如其它细胞核酸或蛋白等其它污染物中将核酸纯化出来时，在核酸是“分离的”或“基本纯化的”。参见F.Ausubel et al.，ed.CurrentProtocols in MolecularBiology，Greene Publishing and Wiley InterScience NewYork(1987)。When the nucleic acid is purified from other cellular components or other contaminants such as other cellular nucleic acids or proteins by standard techniques including alkali/SDS treatment, CsCl banding, column chromatography, agarose gel electrophoresis and other techniques known in the art When the nucleic acid is "isolated" or "substantially purified". See F. Ausubel et al., ed. Current Protocols in Molecular Biology, Greene Publishing and Wiley InterScience New York (1987).

当将其与另一个核酸序列进行功能相连时，则核酸是“可操作连接的”。例如，启动子或增强子当影响序列的转录时则是与编码序列可操作连接的。对于调控序列的转录而言，可操作连接是指被连接的DNA序列在连接两个蛋白编码区是连续的，而且使阅读框也是连续的。对开关序列而言，可操作连接表示可影响开关重组的序列。Nucleic acids are "operably linked" when they are functionally linked to another nucleic acid sequence. For example, a promoter or enhancer is operably linked to a coding sequence when it affects the transcription of the sequence. For the transcription of regulatory sequences, operably linked means that the DNA sequences being linked are contiguous in connecting two protein coding regions, and the reading frames are also contiguous. With respect to switch sequences, operably linked means sequences that can affect switch recombination.

此处所用术语“抑制生长”(例如当指细胞时)是指当与CD38BP，例如抗CD38抗体接触时，和未与CD38BP，例如抗CD38抗体接触时相比较，细胞生长的任何可测定的降低，例如抑制细胞培养物生长的至少约10％、20％、30％、40％、50％、60％、70％、80％、90％、99％或100％。The term "growth inhibition" (e.g. when referring to cells) as used herein refers to any measurable decrease in cell growth when contacted with a CD38BP, e.g., an anti-CD38 antibody, compared to when not contacted with a CD38BP, e.g., an anti-CD38 antibody , such as inhibiting the growth of a cell culture by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 99%, or 100%.

此处所用术语“抑制结合”和“阻断结合”(例如当指抑制/阻断CD38结合物与CD38结合时)可互换使用，均包括部分和完全抑制/阻断。CD38结合物与CD38的结合的抑制/阻断可降低或改变CD38结合物与CD38结合未被抑制或阻断时发生的细胞信号传导的正常水平或类型。抑制和阻断也指当与诸如抗CD38抗体这样的CD38BP接触时，与未与诸如抗CD38抗体这样的CD38BP接触的配体相比较，CD38结合物与CD38的结合亲和性任何可测定的降低，例如抑制CD38结合物与CD38的结合至少约10％、20％、30％、40％、50％、60％、70％、80％、90％、99％或100％。As used herein, the terms "inhibiting binding" and "blocking binding" (eg, when referring to inhibiting/blocking the binding of a CD38 binder to CD38) are used interchangeably and both include partial and complete inhibition/blocking. Inhibition/blocking of the binding of the CD38 binder to CD38 can reduce or alter the normal level or type of cellular signaling that occurs when the binding of the CD38 binder to CD38 is not inhibited or blocked. Inhibition and blocking also refer to any measurable decrease in the binding affinity of a CD38 binder to CD38 when contacted with a CD38BP, such as an anti-CD38 antibody, compared to a ligand that is not contacted with a CD38BP, such as an anti-CD38 antibody , such as inhibiting the binding of a CD38-conjugate to CD38 by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 99%, or 100%.

“靶细胞”是指可被本发明的组分(包括例如，诸如人单克隆抗CD38抗体和/或针对CD38的双特异性或多特异性分子这样的CD38BP)作为靶标的受试者(例如人或动物)中的任何非期望的细胞。在某些技术方案中，靶细胞是表达或过表达CD38的细胞。表达CD38的细胞典型地包括诸如骨髓胸腺细胞、激活的T和B细胞、80％其余的NK细胞和单核细胞、淋巴结生发中心成淋巴细胞、浆B细胞和某些滤泡内细胞、枝状细胞、正常骨髓细胞、特定的前体细胞、50-80％脐带血细胞、红血球以及血小板这样的造血细胞。CD38也可被诸如内脏中的上皮内细胞和固有层淋巴细胞这样的非造血细胞、被脑中的Purkinje细胞和神经原纤维缠结、被前列腺的上皮细胞、胰脏中的β细胞、骨中的破骨细胞、眼中的视网膜细胞和平滑肌和横纹肌中的肌膜所表达。在恶性细胞中，CD38在各种恶性血液疾病，包括但不限定于多发性骨髓瘤、原发和继发浆细胞白血病、B细胞慢性淋巴细胞白血病、B细胞急性淋巴细胞白血病、Waldenstrom巨球蛋白血症、原发性系统性淀粉样变病、套细胞淋巴瘤、前淋巴细胞/髓细胞白血病、急性骨髓白血病、慢性骨髓白血病、滤泡淋巴瘤和NK细胞白血病中也有表达。"Target cell" refers to a subject (e.g. Any undesired cell in a human or animal). In some embodiments, the target cells are cells expressing or overexpressing CD38. Cells expressing CD38 typically include cells such as bone marrow thymocytes, activated T and B cells, 80% of the remaining NK cells and monocytes, lymph node germinal center lymphoblasts, plasma B cells and certain intrafollicular cells, dendritic cells, normal bone marrow cells, specific precursor cells, 50-80% cord blood cells, red blood cells, and hematopoietic cells such as platelets. CD38 is also detected by non-hematopoietic cells such as intraepithelial cells and lamina propria lymphocytes in the gut, by Purkinje cells and neurofibrillary tangles in the brain, by epithelial cells in the prostate, beta cells in the pancreas, bone Expressed by osteoclasts in the eye, retinal cells in the eye, and sarcolemma in smooth and striated muscles. In malignant cells, CD38 is involved in various hematological malignancies, including but not limited to multiple myeloma, primary and secondary plasma cell leukemia, B-cell chronic lymphocytic leukemia, B-cell acute lymphoblastic leukemia, Waldenstrom macroglobulin It is also expressed in leukemia, primary systemic amyloidosis, mantle cell lymphoma, prolymphocytic/myeloid leukemia, acute myeloid leukemia, chronic myeloid leukemia, follicular lymphoma and NK cell leukemia.

此处所用术语“载体”是指可运输与其连接的另一种核酸的核酸分子。一种类型的载体是“质粒”，它是指环状双链DNA环，其中可连接额外的DNA片段。另一种类型的载体是病毒载体，其中额外的DNA片段可连接到病毒基因组中。某些载体可在其所导入的宿主细胞中自主复制(例如具有细菌复制起点的细菌载体和游离的哺乳动物载体)。其它载体(例如非游离哺乳动物载体)可在导入到宿主细胞后整合到宿主细胞的基因组中，然后与宿主基因组一起复制。此外，某些载体可指导与其可操作连接的基因的表达。这类载体在此处被称为“重组表达载体”(或简言之，“表达载体”)。一般来说，应用于重组DNA技术中的表达载体通常是质粒形式。在本说明书中，由于质粒是载体的最普遍的使用形式，因此“质粒”和“载体”可互换使用。但本发明包括其它形式的表达载体，例如病毒载体(比如复制缺陷的逆转录病毒、腺病毒和腺相关病毒，它们可发挥同样的功能。The term "vector" as used herein refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a "plasmid", which refers to a circular double-stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, in which additional DNA segments can be ligated into the viral genome. Certain vectors replicate autonomously in the host cell into which they are introduced (eg, bacterial vectors with a bacterial origin of replication and episomal mammalian vectors). Other vectors (eg, non-episomal mammalian vectors) can integrate into the genome of the host cell after introduction into the host cell and then replicate with the host genome. In addition, certain vectors direct the expression of genes to which they are operably linked. Such vectors are referred to herein as "recombinant expression vectors" (or, simply, "expression vectors"). In general, expression vectors used in recombinant DNA techniques are usually in the form of plasmids. In this specification, "plasmid" and "vector" are used interchangeably since plasmid is the most commonly used form of vector. However, the invention includes other forms of expression vectors, such as viral vectors (such as replication defective retroviruses, adenoviruses and adeno-associated viruses), which serve the same function.

此处所用术语“重组宿主细胞”(或简言之“宿主细胞”)是指导入重组表达载体的细胞。可理解为该术语不仅是指特定的受试细胞，而且也指这种细胞的后代。由于在后代中因突变或环境影响可能发生某些 改变，因此事实上，这种后代可能与母细胞不是相同的，但仍包括在此处所用术语“宿主细胞”的范围中。重组宿主细胞包括例如诸如CHO细胞这样的转染瘤，NS/0细胞和淋巴细胞。The term "recombinant host cell" (or simply "host cell") as used herein refers to a cell into which a recombinant expression vector has been introduced. It is understood that the term refers not only to the particular subject cell, but also to the progeny of such a cell. Such progeny may not in fact be identical to the parent cell due to certain changes that may occur in the progeny due to mutations or environmental influences, but are still included within the scope of the term "host cell" as used herein. Recombinant host cells include, for example, transfectomas such as CHO cells, NS/0 cells and lymphocytes.

术语“调控序列”包括可控制抗体链基因转录或翻译的启动子、增强子和其它表达控制元件(例如多聚腺苷酸信号)。这种调控序列可描述在例如Goeddel，Gene ExpressionTechnology.Methods in Enzymology185，Academic Press，San Diego，Calif.(1990)中。本领域技术人员可设计表达载体，包括根据选择被转化的宿主细胞、所需蛋白的表达水平等因素来选择调控序列。用于哺乳动物宿主细胞表达的调控序列的例子包括可在哺乳动物中指导高水平蛋白表达的病毒元件，例如来源于]细胞巨化病毒(CMV)、猿病毒40(SV40)、腺病毒(例如腺病毒主要晚期启动子(AdMLP))和多瘤病毒的启动子和/或增强子。选择性地，可使用非病毒调控序列，例如泛素启动子β-球蛋白启动子。The term "regulatory sequence" includes promoters, enhancers and other expression control elements (eg, polyadenylation signals) that control the transcription or translation of the antibody chain genes. Such regulatory sequences are described, for example, in Goeddel, Gene Expression Technology. Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990). Those skilled in the art can design expression vectors, including selection of regulatory sequences based on selection of transformed host cells, expression levels of desired proteins and other factors. Examples of regulatory sequences for expression in mammalian host cells include viral elements that direct high-level protein expression in mammals, such as those derived from cytomegalovirus (CMV), simian virus 40 (SV40), adenovirus (e.g. Adenovirus major late promoter (AdMLP)) and polyomavirus promoters and/or enhancers. Alternatively, non-viral regulatory sequences can be used, such as the ubiquitin promoter β-globin promoter.

此处所用术语“受试体”包括任何人和非人动物。术语“非人动物”包括所有脊椎动物，例如哺乳动物和非哺乳动物，例如非人灵长类、羊、狗、牛、鸡、两栖类、爬行类等。The term "subject" as used herein includes any human and non-human animals. The term "non-human animal" includes all vertebrates, such as mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, and the like.

术语“转染”的各种形式包括广泛的各种通常用于将外源DNA导入原核或真核细胞的技术，例如电穿孔、磷酸钙沉淀、DEAE-右旋糖苷转染、脂质体转染等。The various forms of the term "transfection" include a wide variety of techniques commonly used to introduce foreign DNA into prokaryotic or eukaryotic cells, such as electroporation, calcium phosphate precipitation, DEAE-dextran transfection, liposome transfection, etc. Dyeing etc.

此处所用术语“转染瘤”包括表达抗体的重组真核宿主细胞，例如CHO细胞、NS/0细胞、HEK293细胞、植物细胞或真菌，包括酵母细胞。The term "transfectoma" as used herein includes recombinant eukaryotic host cells expressing antibodies, such as CHO cells, NS/0 cells, HEK293 cells, plant cells or fungi, including yeast cells.

术语“非人动物”包括所有脊椎动物，例如，哺乳动物和非哺乳动物，譬如非人灵长类、羊、狗、牛、鸡、两栖类、爬行类等。术语“非人动物”包括所有脊椎动物，例如，哺乳动物和非哺乳动物，譬如非人灵长类、羊、狗、牛、鸡、两栖类、爬行类等。术语“非人动物”包括所有脊椎动物，例如，哺乳动物和非哺乳动物，譬如非人灵长类、羊、狗、牛、鸡、两栖类、爬行类等。The term "non-human animal" includes all vertebrates, eg, mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, and the like. The term "non-human animal" includes all vertebrates, eg, mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, and the like. The term "non-human animal" includes all vertebrates, eg, mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, and the like.

术语“转基因非人动物”是指具有含一个或多个人重链和/或轻链转基因或转染色体的基因组(整合或不整合在动物的天然基因组DNA中)，并可完全表达人抗体的非人动物。例如，转基因鼠可具有人轻链转基因或人重链转染色体，从而当鼠用CD38抗原和/或表达CD38的细胞免疫时可产生人可CD38抗体。人重链转基因可整合在鼠染色体DNA 中，这种情况下的转基因鼠是，例如诸如HCo7或HCo12鼠这样的HuMAb鼠，或人重链转基因可维持在染色体外，这种情况下的转染色体KM鼠在WO02/43478中有所描述。这种转基因和转染色体鼠(此处统称为“转基因鼠”)可通过V-D-J重组和同型体转换产生多种针对给定抗原的人单克隆抗体的同型体(例如IgG、IgA、IgM、IgD和/或IgE)。转基因非人动物也可通过导入编码特定抗体的基因、例如通过将基因与在动物乳汁中表达的基因有效连接的方式来用于产生针对特定抗原的抗体The term "transgenic non-human animal" refers to a non-human animal having a genome containing one or more human heavy and/or light chain transgenes or transchromosomes (integrated or not integrated into the animal's native genomic DNA) and capable of fully expressing human antibodies. human animal. For example, a transgenic mouse can have a human light chain transgene or a human heavy chain transchromosome so that when the mouse is immunized with a CD38 antigen and/or CD38-expressing cells, it can produce human CD38 antibodies. The human heavy chain transgene can be integrated in the mouse chromosomal DNA, in which case the transgenic mouse is, for example, a HuMAb mouse such as HCo7 or HCo12 mice, or the human heavy chain transgene can be maintained extrachromosomally, in which case the transchromosomal KM mice are described in WO02/43478. Such transgenic and transchromosomal mice (collectively referred to herein as "transgenic mice") can produce multiple isotypes (e.g., IgG, IgA, IgM, IgD, and and/or IgE). Transgenic non-human animals can also be used to produce antibodies to specific antigens by introducing a gene encoding the specific antibody, for example, by operably linking the gene to a gene expressed in the milk of the animal

此处所用术语特异性是指诸如抗CD38抗体这样的CD38结合肽识别CD38抗原决定簇、但对CD38的其它部分(包括可被诸如抗CD38抗体这样的其它CD38BPs结合的其它的抗原决定簇)仅具有很小或不可检测的反应性的能力。特异性可通过此处所述的竞争检测法进行相关检测。特异性更具体地可通过任何此处所述抗原决定簇鉴定/描述技术或其在本领域中已知的等效技术来测定。The term specificity as used herein means that a CD38 binding peptide, such as an anti-CD38 antibody, recognizes a CD38 epitope, but only to other parts of CD38 (including other epitopes that can be bound by other CD38BPs such as an anti-CD38 antibody). The ability to have little or no detectable reactivity. Specificity can be correlatively tested by competition assays as described herein. Specificity may more specifically be determined by any of the antigenic determinant identification/characterization techniques described herein or their equivalent known in the art.

不过特异针对特定抗原决定簇的抗体可与某些存在CD38的生物样品中的其它分子发生交叉反应。更典型地，诸如抗CD38抗体这样的CD38BP，可与来自其它物种的CD38同源物交叉反应。在其中一种或全部两种情况下，典型地，这种交叉反应抗体是针对与人CD38相关的结构和/或环境因子进行选择的。However, antibodies specific for a particular epitope may cross-react with other molecules in some biological samples where CD38 is present. More typically, CD38BPs, such as anti-CD38 antibodies, cross-react with CD38 homologues from other species. In either or both cases, typically such cross-reactive antibodies are selected against structural and/or environmental factors associated with human CD38.

此处所用术语选择性是指诸如抗CD38抗体这样的CD38BP对特定的区域、靶、或者肽的优先结合；典型地，相对于一个或多个其它生物分子、结构、细胞、组织等，是CD38上的区域或抗原决定簇。在一个技术方案中，本发明中的诸如抗CD38抗体这样的CD38BP是针对直肠癌细胞环境中的CD38部分进行选择的(即，抗CD38抗体选择性地结合到CD38部分，超过与直肠癌细胞其它组分的结合)。The term selectivity as used herein refers to the preferential binding of a CD38BP, such as an anti-CD38 antibody, to a specific region, target, or peptide; typically, CD38 relative to one or more other biomolecules, structures, cells, tissues, etc. region or antigenic determinant. In one embodiment, a CD38BP such as an anti-CD38 antibody of the invention is selected against a portion of CD38 in the environment of colorectal cancer cells (i.e., an anti-CD38 antibody binds selectively to a portion of CD38 over other parts of colorectal cancer cells). combination of components).

本发明的CD38BPs典型地以一种至少是充分分离的形式来使用并提供的。充分分离的分子是在组分中为主要种类的分子，在上述的组分中含该分子与其所属的一类分子(即，它在组分中至少占约50％的分子类型，典型地在组分中至少占约70％、至少约80％、至少占约85％、至少约90％、至少约95％、或更多的分子种类，例如肽(譬如，对所有存在的肽种类中，组分对CD38BP具有至少约98％、98％或99％的同质性))。The CD38BPs of the invention are typically used and provided in an at least substantially isolated form. A well-separated molecule is one that is the predominant species in the fraction in which it is contained along with the class of molecules to which it belongs (i.e., a molecular type that makes up at least about 50% of the fraction, typically in At least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or more molecular species, such as peptides (e.g., for all peptide species present, The components have at least about 98%, 98% or 99% homogeneity to CD38BP)).

分离的分子是指不与诸如产生CD38BP的细胞或动物中所含有的非CD38结合生物分子(或可干扰本发明的CD38BP结合和/活性的CD38结合分子)这样的任何外源及非必需的生理因子在显著性上(例如大于约1％、大于约2％、大于约3％、或大于约5％)相关联的分子。分离的分子也可指任何经过人工干预(无论自动、手工还是二者皆有)的纯化阶段所获得的分子。在本发明提供的多种组分中，例如在含有一个或多个药学上可接受的载体的组分中(例如在含有大量药学上可接受的载体、稳定剂和/或防腐剂的组分的情况下)，CD38BP可根据组分中所有分子种类的数目而以相对较小的量存在。在某些情况下，诸如BSA这样的附加的肽，也可与先前纯化的CD38BP包括在这样的组分中。不过，如果这种组分的附加组成对于有意使用CD38BP是可以接受的，那么这种组分仍可描述为包含分离的CD38BP。An isolated molecule is one that is free from any exogenous and non-essential physiological molecules such as non-CD38-binding biomolecules contained in CD38BP-producing cells or animals (or CD38-binding molecules that can interfere with CD38BP binding and/activity of the present invention). Molecules for which factors are associated in a significant degree (eg, greater than about 1%, greater than about 2%, greater than about 3%, or greater than about 5%). An isolated molecule may also refer to any molecule obtained through a purification stage that has undergone human intervention, whether automated, manual or both. In the various compositions provided by the invention, for example, in the composition that contains one or more pharmaceutically acceptable carriers (for example, in the composition that contains a large amount of pharmaceutically acceptable carriers, stabilizers and/or preservatives case), CD38BP may be present in relatively small amounts depending on the number of all molecular species in the component. In some cases, additional peptides, such as BSA, can also be included in such fractions with previously purified CD38BP. However, such components can still be described as comprising isolated CD38BP if the additional composition of such components is acceptable for the intended use of CD38BP.

典型地，本发明的CD38BPs实质上不含有诸如具有不同抗原特异性的CD38BPs这样的其它CD38BPs。但本发明也提供了含有多种具有不同特异性和特征的CD38BPs的组分(例如，本发明提供了具有不同特异性和/或选择性特征的CD38BPs“鸡尾酒”)。Typically, the CD38BPs of the invention are substantially free of other CD38BPs such as CD38BPs with different antigenic specificities. However, the invention also provides compositions comprising multiple CD38BPs with different specificities and characteristics (eg, the invention provides "cocktails" of CD38BPs with different specificity and/or selectivity characteristics).

“治疗”是指给药有效剂量的本发明的具有治疗活性的化合物，以达到减轻、减弱或根除(治愈)症状或疾病状态的目的。"Treatment" refers to the administration of an effective dose of a therapeutically active compound of the present invention for the purpose of alleviating, weakening or eradicating (curing) a symptom or disease state.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：2的序列所组成的V_L的CD38BP。In one technical solution, the present invention provides CD38BP containing a _VL substantially consisting of the sequence of SEQ ID No:2.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：6的序列所组成的V_H的CD38BP。In one technical solution, the present invention provides CD38BP comprising a _VH substantially consisting of the sequence of SEQ ID No:6.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：2的序列所组成的V_L和实质上由SEQ ID No：6的序列所组成的V_H的CD38BP。In one technical solution, the present invention provides CD38BP comprising a V _L substantially consisting of the sequence of SEQ ID No: 2 and a V _H substantially consisting of the sequence of SEQ ID No: 6.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：3的序列所组成的V_LCDR1的CD38BP。In one technical solution, the present invention provides CD38BP containing V _L CDR1 substantially consisting of the sequence of SEQ ID No:3.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：4的序列所组成的V_LCDR2的CD38BP。In one technical solution, the present invention provides CD38BP containing V _L CDR2 substantially consisting of the sequence of SEQ ID No:4.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：5的序列所组成的V_LCDR3的CD38BP。In one technical solution, the present invention provides CD38BP comprising V _L CDR3 substantially consisting of the sequence of SEQ ID No:5.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：8的 序列所组成的V_HCDR1的CD38BP。In one technical solution, the present invention provides CD38BP comprising a _VH CDR1 substantially consisting of the sequence of SEQ ID No:8.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：9的序列所组成的V_HCDR2的CD38BP。In one technical solution, the present invention provides CD38BP comprising a _VH CDR2 substantially consisting of the sequence of SEQ ID No:9.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：10的序列所组成的V_HCDR3的CD38BP。In one technical solution, the present invention provides CD38BP comprising a _VH CDR3 substantially consisting of the sequence of SEQ ID No:10.

在一个技术方案中，本发明提供了含有实质上分别由SEQ ID No：3、SEQ ID No：4和SEQ ID No：5的序列所组成的V_L CDRs(V_L CDR1、CDR2和CDR3)的CD38BP。In one technical solution, the present invention provides V _L CDRs (V _L CDR1, CDR2 and CDR3) comprising substantially the sequences of SEQ ID No: 3, SEQ ID No: 4 and SEQ ID No: 5 respectively CD38BP.

在一个技术方案中，本发明提供了含有实质上分别由SEQ ID No：8、SEQ ID No：9和SEQ ID No：10的序列所组成的V_H CDRs(V_H CDR1、CDR2和CDR3)的CD38BP。In one technical solution, the present invention provides _VH CDRs ( _VH CDR1, CDR2 and CDR3) comprising substantially the sequences of SEQ ID No: 8, SEQ ID No: 9 and SEQ ID No: 10, respectively. CD38BP.

在一个技术方案中，本发明提供了CD38BP，它包含In one technical solution, the present invention provides CD38BP, which comprises

(a)三种V_L CDRs，它们实质上分别由SEQ ID No：3、SEQ ID No：4和SEQ ID No：5所组成，并在CD38BP中彼此紧邻(例如，在野生型抗CD38抗体中靠近V_L CDRs的间隔区)，以及(a) Three V _L CDRs, which consist essentially of SEQ ID No: 3, SEQ ID No: 4, and SEQ ID No: 5, respectively, and are located next to each other in CD38BP (for example, in a wild-type anti-CD38 antibody spacers near the V _L CDRs), and

(b)三种V_H CDRs，它们实质上分别由SEQ ID No：8、SEQ ID No：9和SEQ ID No：10所组成，并在CD38BP中彼此紧邻(例如，在野生型抗CD38抗体中靠近V_H CDRs的间隔区)。(b) three _VH CDRs, which consist essentially of SEQ ID No: 8, SEQ ID No: 9 and SEQ ID No: 10, respectively, and are located next to each other in CD38BP (for example, in a wild-type anti-CD38 antibody spacers near the _VH CDRs).

在一个进一步的技术方案中，本发明提供了在CD38BP的V_L区和V_H区之间含有柔性连接物的CD38BP。在另一个进一步的技术方案中，本发明提供了一种CD38BP，其中V_L和V_H区位于免疫球蛋白折叠蛋白的不同链上，并以V_L CDR1、CDR2、CDR3及V_H CDR1、CDR2和CDR3这样的顺序定向连接，从而可与CD38上的抗原决定簇选择性和/或特异性结合。在另一个进一步的技术方案中，本发明提供了含有两组可变结构域(在相联的不同链上联结的V_L和V_H结构域组)的CD38BP，从而使CD38BP含有两个相同的抗原决定簇结合位点。In a further technical solution, the present invention provides CD38BP containing a flexible linker between the _VL and _VH regions of CD38BP. In another further technical solution, the present invention provides a CD38BP, wherein the V _L and V _H regions are located on different chains of the immunoglobulin folded protein, and V _L CDR1, CDR2, CDR3 and V _H CDR1, CDR2 Directly linked with the sequence of CDR3, so as to selectively and/or specifically bind to the antigenic determinant on CD38. In another further technical solution, the present invention provides a CD38BP containing two sets of variable domains ( _VL and _VH domain sets linked on different chains) so that the CD38BP contains two identical Antigen determinant binding site.

预期在本段中所述的任何这种CD38BPs，至少在部分上，与具有含SEQ ID No：2序列的V_L区和含SEQ ID No：7序列的V_H区的抗体，具有相似的抗原决定簇特异性、选择性和其它特征，因此，可用于治疗多发性骨髓瘤。It is contemplated that any such CD38BPs described in this paragraph, at least in part, have similar antigenicity to an antibody having a _VL region comprising the sequence of SEQ ID No: 2 and a _VH region comprising the sequence of SEQ ID No: 7 Determinant specificity, selectivity and other features, therefore, can be used in the treatment of multiple myeloma.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：12的序列所组成的V_L的CD38BP。In one technical solution, the present invention provides CD38BP comprising a _VL substantially consisting of the sequence of SEQ ID No: 12.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：17的序列所组成的V_H的CD38BP。In one technical solution, the present invention provides CD38BP comprising a _VH substantially consisting of the sequence of SEQ ID No: 17.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：12的序列所组成的V_L和实质上由SEQ ID No：17的序列所组成的V_H的CD38BP。In one technical solution, the present invention provides CD38BP comprising a V _L substantially consisting of the sequence of SEQ ID No: 12 and a V _H substantially consisting of the sequence of SEQ ID No: 17.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：13的序列所组成的V_LCDR1的CD38BP。In one technical solution, the present invention provides CD38BP containing V _L CDR1 substantially consisting of the sequence of SEQ ID No: 13.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：14的序列所组成的V_LCDR2的CD38BP。In one technical solution, the present invention provides CD38BP containing V _L CDR2 substantially consisting of the sequence of SEQ ID No: 14.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：15的序列所组成的V_LCDR3的CD38BP。In one technical solution, the present invention provides CD38BP containing V _L CDR3 substantially consisting of the sequence of SEQ ID No: 15.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：18的序列所组成的V_HCDR1的CD38BP。In one technical solution, the present invention provides CD38BP comprising a _VH CDR1 substantially consisting of the sequence of SEQ ID No: 18.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：19的序列所组成的V_HCDR2的CD38BP。In one technical solution, the present invention provides CD38BP comprising a _VH CDR2 substantially consisting of the sequence of SEQ ID No: 19.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：20的序列所组成的V_HCDR3的CD38BP。In one technical solution, the present invention provides CD38BP comprising a _VH CDR3 substantially consisting of the sequence of SEQ ID No:20.

在一个技术方案中，本发明提供了含有实质上分别由SEQ ID No：13、SEQ ID No：14和SEQ ID No：15的序列所组成的V_L CDRs(V_L CDR1、CDR2和CDR3)的CD38BP。In one technical solution, the present invention provides V _L CDRs (V _L CDR1, CDR2 and CDR3) comprising substantially the sequences of SEQ ID No: 13, SEQ ID No: 14 and SEQ ID No: 15 respectively CD38BP.

在一个技术方案中，本发明提供了含有实质上分别由SEQ ID No：18、SEQ ID No：19和SEQ ID No：20的序列所组成的V_H CDRs(V_H CDR1、CDR2和CDR3)的CD38BP。In one technical solution, the present invention provides _VH CDRs ( _VH CDR1, CDR2 and CDR3) comprising substantially the sequences of SEQ ID No: 18, SEQ ID No: 19 and SEQ ID No: 20 respectively CD38BP.

在一个技术方案中，本发明提供了CD38BP，它包含In one technical solution, the present invention provides CD38BP, which comprises

(a)三种V_L CDRs，它们实质上分别由SEQ ID No：13、SEQ ID No：14和SEQ ID No：15所组成，并在CD3 8BP中彼此紧邻(例如，在野生型抗CD3 8抗体中靠近V_L CDRs的间隔区)，以及(a) Three V _L CDRs, which consist essentially of SEQ ID No: 13, SEQ ID No: 14, and SEQ ID No: 15, respectively, and are located next to each other in the CD3 8BP (for example, in wild-type anti-CD3 8BP spacers near the V _L CDRs in the antibody), and

(b)三种V_H CDRs，它们实质上分别由SEQ ID No：18、SEQ ID No：19和SEQ ID No：20所组成，并在CD3 8BP中彼此紧邻(例如，在野生型抗CD38抗体中靠近V_H CDRs的间隔区)。(b) three _VH CDRs, which consist essentially of SEQ ID No: 18, SEQ ID No: 19 and SEQ ID No: 20, respectively, and are located next to each other in the CD3 8BP (for example, in a wild-type anti-CD38 antibody spacers near the _VH CDRs).

在一个进一步的技术方案中，本发明提供了在CD38BP的V_L区和V_H区之间含有柔性连接物的CD38BP。在另一个进一步的技术方案中， 本发明提供了一种CD38BP，其中V_L和V_H区位于免疫球蛋白折叠蛋白的不同链上，并以V_L CDR1、CDR2、CDR3及V_H CDR1、CDR2和CDR3这样的顺序定向连接，从而可与CD38上的抗原决定簇选择性和/或特异性结合。在另一个进一步的技术方案中，本发明提供了含有两组可变结构域(在相联的不同链上联结的V_L和V_H结构域组)的CD38BP，从而使CD38BP含有两个相同的抗原决定簇结合位点。In a further technical solution, the present invention provides CD38BP containing a flexible linker between the _VL and _VH regions of CD38BP. In another further technical solution, the present invention provides a CD38BP, wherein the V _L and V _H regions are located on different chains of the immunoglobulin folded protein, and V _L CDR1, CDR2, CDR3 and V _H CDR1, CDR2 Directly linked with the sequence of CDR3, so as to selectively and/or specifically bind to the antigenic determinant on CD38. In another further technical solution, the present invention provides a CD38BP containing two sets of variable domains ( _VL and _VH domain sets linked on different chains) so that the CD38BP contains two identical Antigen determinant binding site.

预期在本段中所述的任何这种CD38BPs，至少在部分上，与具有含SEQ ID No：12序列的V_L区和含SEQ ID No：17序列的V_H区的抗体，具有相似的抗原决定簇特异性、选择性和其它特征，因此，可用于治疗多发性骨髓瘤。Any such CD38BPs described in this paragraph are expected, at least in part, to have similar antigenicity to an antibody having a _VL region comprising the sequence of SEQ ID No: 12 and a _VH region comprising the sequence of SEQ ID No: 17 Determinant specificity, selectivity and other features, therefore, can be used in the treatment of multiple myeloma.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：22的序列所组成的V_L的CD38BP。In one technical solution, the present invention provides a CD38BP comprising a _VL substantially consisting of the sequence of SEQ ID No: 22.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：27的序列所组成的V_H的CD38BP。In one technical solution, the present invention provides CD38BP comprising a _VH substantially consisting of the sequence of SEQ ID No: 27.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：22的序列所组成的V_L和实质上由SEQ ID No：27的序列所组成的V_H的CD38BP。In one technical solution, the present invention provides a CD38BP comprising a V _L substantially consisting of the sequence of SEQ ID No: 22 and a V _H substantially consisting of the sequence of SEQ ID No: 27.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：23的序列所组成的V_LCDR1的CD38BP。In one technical solution, the present invention provides CD38BP containing V _L CDR1 substantially consisting of the sequence of SEQ ID No: 23.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：24的序列所组成的V_LCDR2的CD38BP。In one technical solution, the present invention provides CD38BP containing V _L CDR2 substantially consisting of the sequence of SEQ ID No: 24.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：25的序列所组成的V_LCDR3的CD38BP。In one technical solution, the present invention provides CD38BP comprising V _L CDR3 substantially consisting of the sequence of SEQ ID No: 25.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：28的序列所组成的V_HCDR1的CD38BP。In one technical solution, the present invention provides CD38BP comprising a _VH CDR1 substantially consisting of the sequence of SEQ ID No: 28.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：29的序列所组成的V_HCDR2的CD38BP。In one technical solution, the present invention provides CD38BP comprising a _VH CDR2 substantially consisting of the sequence of SEQ ID No: 29.

在一个技术方案中，本发明提供了含有实质上由SEQ ID No：30的序列所组成的V_HCDR3的CD38BP。In one technical solution, the present invention provides CD38BP comprising a _VH CDR3 substantially consisting of the sequence of SEQ ID No: 30.

在一个技术方案中，本发明提供了含有实质上分别由SEQ ID No：23、SEQ ID No：24和SEQ ID No：25的序列所组成的V_L CDRs(V_L CDR1、CDR2和CDR3)的CD38BP。In one technical solution, the present invention provides V _L CDRs (V _L CDR1, CDR2 and CDR3) comprising substantially the sequences of SEQ ID No: 23, SEQ ID No: 24 and SEQ ID No: 25 respectively CD38BP.

在一个技术方案中，本发明提供了含有实质上分别由SEQ ID No：28、SEQ ID No：29和SEQ ID No：30的序列所组成的V_H CDRs(V_H CDR1、CDR2和CDR3)的CD38BP。In one technical solution, the present invention provides _VH CDRs ( _VH CDR1, CDR2 and CDR3) comprising substantially the sequences of SEQ ID No: 28, SEQ ID No: 29 and SEQ ID No: 30 respectively CD38BP.

在一个技术方案中，本发明提供了CD38BP，它包含In one technical solution, the present invention provides CD38BP, which comprises

(a)三种V_L CDRs，它们实质上分别由SEQ ID No：23、SEQ ID No：24和SEQ ID No：25所组成，并在CD38BP中彼此紧邻(例如，在野生型抗CD38抗体中靠近V_L CDRs的间隔区)，以及(a) Three V _L CDRs, which consist essentially of SEQ ID No: 23, SEQ ID No: 24, and SEQ ID No: 25, respectively, and are located next to each other in CD38BP (for example, in a wild-type anti-CD38 antibody spacers near the V _L CDRs), and

(b)三种V_H CDRs，它们实质上分别由SEQ ID No：28、SEQ ID No：29和SEQ ID No：30所组成，并在CD38BP中彼此紧邻(例如，在野生型抗CD38抗体中靠近V_H CDRs的间隔区)。(b) three _VH CDRs, which consist essentially of SEQ ID No: 28, SEQ ID No: 29 and SEQ ID No: 30, respectively, and are located next to each other in CD38BP (for example, in a wild-type anti-CD38 antibody spacers near the _VH CDRs).

在一个进一步的技术方案中，本发明提供了在CD38BP的V_L区和V_H区之间含有柔性连接物的CD38BP。在另一个进一步的技术方案中，本发明提供了一种CD38BP，其中V_L和V_H区位于免疫球蛋白折叠蛋白的不同链上，并以V_L CDR1、CDR2、CDR3及V_H CDR1、CDR2和CDR3这样的顺序定向连接，从而可与CD38上的抗原决定簇选择性和/或特异性结合。在另一个进一步的技术方案中，本发明提供了含有两组可变结构域(在相联的不同链上联结的V_L和V_H结构域组)的CD38BP，从而使CD38BP含有两个相同的抗原决定簇结合位点。In a further technical solution, the present invention provides CD38BP containing a flexible linker between the _VL and _VH regions of CD38BP. In another further technical solution, the present invention provides a CD38BP, wherein the V _L and V _H regions are located on different chains of the immunoglobulin folded protein, and V _L CDR1, CDR2, CDR3 and V _H CDR1, CDR2 Directly linked with the sequence of CDR3, so as to selectively and/or specifically bind to the antigenic determinant on CD38. In another further technical solution, the present invention provides a CD38BP containing two sets of variable domains ( _VL and _VH domain sets linked on different chains) so that the CD38BP contains two identical Antigen determinant binding site.

预期在本段中所述的任何这种CD38BPs，至少在部分上，与具有含SEQ ID No：22序列的V_L区和含SEQ ID No：27序列的V_H区的抗体，具有相似的抗原决定簇特异性、选择性和其它特征，因此，可用于治疗多发性骨髓瘤。It is contemplated that any such CD38BPs described in this paragraph, at least in part, have similar antigenicity to an antibody having a _VL region comprising the sequence of SEQ ID No: 22 and a _VH region comprising the sequence of SEQ ID No: 27 Determinant specificity, selectivity and other features, therefore, can be used in the treatment of multiple myeloma.

在一个技术方案中，本发明提供了含有实质上由如SEQ ID No：3或SEQ ID No：13或SEQ ID No：23所述的序列组成的V_L CDR1的CD38BP，其中N末端残基和/或1、2或3个C末端氨基酸残基是缺失的。In one technical solution, the present invention provides a CD38BP containing V _L CDR1 substantially consisting of the sequence described in SEQ ID No: 3 or SEQ ID No: 13 or SEQ ID No: 23, wherein the N-terminal residues and /or 1, 2 or 3 C-terminal amino acid residues are deleted.

在一个技术方案中，本发明提供了含有实质上由如SEQ ID No：4或SEQ ID No：14或SEQ ID No：24所述的序列组成的V_L CDR2的CD38BP，其中N末端残基和/或1、2或3个C末端残基是缺失的。In one technical solution, the present invention provides a CD38BP containing V _L CDR2 substantially consisting of the sequence described in SEQ ID No: 4 or SEQ ID No: 14 or SEQ ID No: 24, wherein the N-terminal residues and /or 1, 2 or 3 C-terminal residues are deleted.

在一个技术方案中，本发明提供了含有实质上由如SEQ ID No：5或SEQ ID No：15或SEQ ID No：25所述的序列组成的V_L CDR3的CD38BP，其中N末端残基和/或1、2、3或4个C末端残基是缺失的。In one technical solution, the present invention provides a CD38BP containing V _L CDR3 substantially consisting of the sequence described in SEQ ID No: 5 or SEQ ID No: 15 or SEQ ID No: 25, wherein the N-terminal residues and /or 1, 2, 3 or 4 C-terminal residues are deleted.

在一个技术方案中，本发明提供了含有实质上由如SEQ ID No：8或SEQ ID No：18或SEQ ID No：28所述的序列组成的V_H CDR1的CD38BP，其中1、2、3或4个N末端残基和/或1、2、3或4个C末端残基是缺失的。In one technical solution, the present invention provides CD38BP containing _VH CDR1 substantially consisting of the sequence described in SEQ ID No: 8 or SEQ ID No: 18 or SEQ ID No: 28, wherein 1, 2, 3 Either 4 N-terminal residues and/or 1 , 2, 3 or 4 C-terminal residues are deleted.

在一个技术方案中，本发明提供了含有实质上由如SEQ ID No：9或SEQ ID No：19或SEQ ID No：29所述的序列组成的V_H CDR2的CD38BP，其中1、2、3、4或5个N末端氨基酸和/或1、2、3、4、5或6个C末端氨基酸是缺失的。In one technical solution, the present invention provides CD38BP containing _VH CDR2 substantially consisting of the sequence described in SEQ ID No: 9 or SEQ ID No: 19 or SEQ ID No: 29, wherein 1, 2, 3 , 4 or 5 N-terminal amino acids and/or 1 , 2, 3, 4, 5 or 6 C-terminal amino acids are deleted.

在一个技术方案中，本发明提供了含有实质上由如SEQ ID No：10或SEQ ID No：20或SEQ ID No：30所述的序列组成的V_H CDR3的CD38BP，其中N末端1、2或3个氨基酸残基和/或C末端1、2、3或4个氨基酸残基是缺失的。In one technical solution, the present invention provides a CD38BP containing _VH CDR3 substantially consisting of the sequence described in SEQ ID No: 10 or SEQ ID No: 20 or SEQ ID No: 30, wherein N-terminal 1, 2 or 3 amino acid residues and/or the C-terminal 1, 2, 3 or 4 amino acid residues are deleted.

本发明也提供其中这些“剪切的”CDR序列彼此间和/或其它此处所述CDR序列组合在一起的CD38BPs。The invention also provides CD38BPs in which these "spliced" CDR sequences are combined with each other and/or other CDR sequences described herein.

在一个技术方案中，本发明提供CD38BP，包含In one technical solution, the present invention provides CD38BP, comprising

(a)三种V_L CDRs，它们实质上分别由SEQ ID No：3、SEQ ID No：4和SEQ ID No：5所组成，并在CD38BP中彼此紧邻(例如，在野生型抗CD38抗体中靠近V_L CDRs的间隔区)，以及(a) Three V _L CDRs, which consist essentially of SEQ ID No: 3, SEQ ID No: 4, and SEQ ID No: 5, respectively, and are located next to each other in CD38BP (for example, in a wild-type anti-CD38 antibody spacers near the V _L CDRs), and

(b)三种V_H CDRs，它们实质上分别由SEQ ID No：8、SEQ ID No：9和SEQ ID No：10所组成，并在CD38BP中彼此紧邻(例如，在野生型抗CD38抗体中靠近V_H CDRs的间隔区)。(b) three _VH CDRs, which consist essentially of SEQ ID No: 8, SEQ ID No: 9 and SEQ ID No: 10, respectively, and are located next to each other in CD38BP (for example, in a wild-type anti-CD38 antibody spacers near the _VH CDRs).

在一个进一步的技术方案中，本发明提供了在CD38BP的V_L区和V_H区之间含有柔性连接物的CD38BP。在另一个进一步的技术方案中，本发明提供了一种CD38BP，其中V_L和V_H区位于免疫球蛋白折叠蛋白的不同链上，并以V_L CDR1、CDR2、CDR3及V_H CDR1、CDR2和CDR3这样的顺序定向连接，从而可与CD38上的抗原决定簇选择性和/或特异性结合。在另一个进一步的技术方案中，本发明提供了含有两组可变结构域(在相联的不同链上联结的V_L和V_H结构域组)的CD38BP，从而使CD38BP含有两个相同的抗原决定簇结合位点。预期在本段中所述的任何这种CD38BPs，至少在部分上，与具有含SEQ ID No：2序列的V_L 区和含SEQ ID No：7序列的V_H区的抗体，具有相似的抗原决定簇特异性、选择性和其它特征。In a further technical solution, the present invention provides CD38BP containing a flexible linker between the _VL and _VH regions of CD38BP. In another further technical solution, the present invention provides a CD38BP, wherein the V _L and V _H regions are located on different chains of the immunoglobulin folded protein, and V _L CDR1, CDR2, CDR3 and V _H CDR1, CDR2 Directly linked with the sequence of CDR3, so as to selectively and/or specifically bind to the antigenic determinant on CD38. In another further technical solution, the present invention provides a CD38BP containing two sets of variable domains ( _VL and _VH domain sets linked on different chains) so that the CD38BP contains two identical Antigen determinant binding site. It is contemplated that any such CD38BPs described in this paragraph, at least in part, have similar antigenicity to an antibody having a _VL region comprising the sequence of SEQ ID No: 2 and a _VH region comprising the sequence of SEQ ID No: 7 Determinant specificity, selectivity and other characteristics.

在一个技术方案中，本发明提供CD38BP，包含In one technical solution, the present invention provides CD38BP, comprising

(a)三种V_L CDRs，它们实质上分别由SEQ ID No：13、SEQ ID No：14和SEQ ID No：15所组成，并在CD38BP中彼此紧邻(例如，在野生型抗CD38抗体中靠近V_L CDRs的间隔区)，以及(a) Three V _L CDRs, which consist essentially of SEQ ID No: 13, SEQ ID No: 14, and SEQ ID No: 15, respectively, and are located next to each other in CD38BP (for example, in a wild-type anti-CD38 antibody spacers near the V _L CDRs), and

(b)三种V_H CDRs，它们实质上分别由SEQ ID No：18、SEQ ID No：19和SEQ ID No：20所组成，并在CD38BP中彼此紧邻(例如，在野生型抗CD38抗体中靠近V_H CDRs的间隔区)。(b) three _VH CDRs, which consist essentially of SEQ ID No: 18, SEQ ID No: 19 and SEQ ID No: 20, respectively, and are located next to each other in CD38BP (for example, in a wild-type anti-CD38 antibody spacers near the _VH CDRs).

在一个进一步的技术方案中，本发明提供了在CD38BP的V_L区和V_H区之间含有柔性连接物的CD38BP。In a further technical solution, the present invention provides CD38BP containing a flexible linker between the _VL and _VH regions of CD38BP.

在另一个进一步的技术方案中，本发明提供了一种CD38BP，其中V_L和V_H区位于免疫球蛋白折叠蛋白的不同链上，并以V_L CDR1、CDR2、CDR3及V_H CDR1、CDR2和CDR3这样的顺序定向连接，从而可与CD38上的抗原决定簇选择性和/或特异性结合。在另一个进一步的技术方案中，本发明提供了含有两组可变结构域(在相联的不同链上联结的V_L和V_H结构域组)的CD38BP，从而使CD38BP含有两个相同的抗原决定簇结合位点。预期在本段中所述的任何这种CD38BPs，至少在部分上，与具有含SEQ ID No：12序列的V_L区和含SEQ ID No：17序列的V_H区的抗体，具有相似的抗原决定簇特异性、选择性和其它特征。In another further technical solution, the present invention provides a CD38BP, wherein the V _L and V _H regions are located on different chains of the immunoglobulin folded protein, and V _L CDR1, CDR2, CDR3 and V _H CDR1, CDR2 Directly linked with the sequence of CDR3, so as to selectively and/or specifically bind to the antigenic determinant on CD38. In another further technical solution, the present invention provides a CD38BP containing two sets of variable domains ( _VL and _VH domain sets linked on different chains) so that the CD38BP contains two identical Antigen determinant binding site. Any such CD38BPs described in this paragraph are expected, at least in part, to have similar antigenicity to an antibody having a _VL region comprising the sequence of SEQ ID No: 12 and a _VH region comprising the sequence of SEQ ID No: 17 Determinant specificity, selectivity and other characteristics.

在一个技术方案中，本发明提供CD38BP，包含In one technical solution, the present invention provides CD38BP, comprising

(a)三种V_L CDRs，它们实质上分别由SEQ ID No：23、SEQ ID No：24和SEQ ID No：25所组成，并在CD38BP中彼此紧邻(例如，在野生型抗CD38抗体中靠近V_L CDRs的间隔区)，以及(a) Three V _L CDRs, which consist essentially of SEQ ID No: 23, SEQ ID No: 24, and SEQ ID No: 25, respectively, and are located next to each other in CD38BP (for example, in a wild-type anti-CD38 antibody spacers near the V _L CDRs), and

(b)三种V_H CDRs，它们实质上分别由SEQ ID No：28、SEQ ID No：29和SEQ ID No：30所组成，并在CD38BP中彼此紧邻(例如，在野生型抗CD38抗体中靠近V_H CDRs的间隔区)。(b) three _VH CDRs, which consist essentially of SEQ ID No: 28, SEQ ID No: 29 and SEQ ID No: 30, respectively, and are located next to each other in CD38BP (for example, in a wild-type anti-CD38 antibody spacers near the _VH CDRs).

在一个进一步的技术方案中，本发明提供了在CD38BP的V_L区和V_H区之间含有柔性连接物的CD38BP。In a further technical solution, the present invention provides CD38BP containing a flexible linker between the _VL and _VH regions of CD38BP.

在另一个进一步的技术方案中，本发明提供了一种CD38BP，其中V_L和V_H区位于免疫球蛋白折叠蛋白的不同链上，并以V_L CDR1、CDR2、CDR3及V_H CDR1、CDR2和CDR3这样的顺序定向连接，从而可与CD38上的抗原决定簇选择性和/或特异性结合。在另一个进一步的技术方案中，本发明提供了含有两组可变结构域(在相联的不同链上联 结的V_L和V_H结构域组)的CD38BP，从而使CD3 8BP含有两个相同的抗原决定簇结合位点。预期在本段中所述的任何这种CD38BPs，至少在部分上，与具有含SEQ ID No：22序列的V_L区和含SEQ ID No：27序列的V_H区的抗体，具有相似的抗原决定簇特异性、选择性和其它特征。In another further technical solution, the present invention provides a CD38BP, wherein the V _L and V _H regions are located on different chains of the immunoglobulin folded protein, and V _L CDR1, CDR2, CDR3 and V _H CDR1, CDR2 Directly linked with the sequence of CDR3, so as to selectively and/or specifically bind to the antigenic determinant on CD38. In another further technical solution, the present invention provides a CD38BP containing two sets of variable domains ( _VL and _VH domain sets linked on different chains) so that the CD3 8BP contains two identical the antigenic determinant binding site. It is contemplated that any such CD38BPs described in this paragraph, at least in part, have similar antigenicity to an antibody having a _VL region comprising the sequence of SEQ ID No: 22 and a _VH region comprising the sequence of SEQ ID No: 27 Determinant specificity, selectivity and other characteristics.

本发明也提供含实施例中抗体的V_L区、V_H区或一个或多个CDRs的功能变异体的CD38BPs。CD38BP所用的V_L、V_H或CDR的功能变异体仍允许CD38BP至少保持母抗体中亲和性/强度和/或特异性/选择性的实质部分(至少约50％、60％、70％、80％、90％、95％或更高)，在某些情况下，这种CD38BP可以具有比母抗体更高的亲和性、选择性和/或特异性。The present invention also provides CD38BPs comprising functional variants of the _VL region, _VH region or one or more CDRs of the antibodies of the embodiments. Functional variants of the _VL , _VH or CDRs used by CD38BP still allow CD38BP to retain at least a substantial portion of the affinity/strength and/or specificity/selectivity of the parent antibody (at least about 50%, 60%, 70%, 80%, 90%, 95% or higher), in some cases, this CD38BP may have higher affinity, selectivity and/or specificity than the parent antibody.

在一个技术方案中，本发明提供含有可变的V_L的CD38BP，该V_L 实质上由与如SEQID No：2或SEQ ID No：12或SEQ ID No：22所述的序列具有约50％，譬如至少60％、例如至少约70％、譬如至少约75％、例如至少约80％、譬如至少约85％、例如至少约90％、譬如至少95％约氨基酸序列同一性的序列所组成，其中CD38BP至少具有抗体抗原决定簇结合特征的实质部分(至少约50％、60％、70％、80％、90％、95％或更多)，该抗体分别具有SEQ ID No：2或SEQ ID No：12或SEQ ID No：22的可变V_L序列，譬如分别具有SEQ ID No：2的V_L序列和SEQID No：7的V_H序列的抗体，及分别具有SEQ ID No：12的V_L序列和SEQ IDNo：17的V_H序列的抗体，和分别具有SEQ ID No：22的V_L序列和SEQID No：27的V_H序列的抗体。In one technical solution, the present invention provides _CD38BP containing a variable V _L , which essentially consists of about 50% of the sequence described in SEQ ID No: 2 or SEQ ID No: 12 or SEQ ID No: 22 , such as at least 60%, such as at least about 70%, such as at least about 75%, such as at least about 80%, such as at least about 85%, such as at least about 90%, such as at least 95% amino acid sequence identity. wherein the CD38BP has at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the epitope binding characteristics of an antibody having SEQ ID No: 2 or SEQ ID No: 12 or the variable V _L sequence of SEQ ID No: 22, such as an antibody having a V _L sequence of SEQ ID No: 2 and a V _H sequence of SEQ ID No: 7, respectively, and a V sequence of SEQ ID No: 12, respectively An antibody having the _L sequence and the _VH sequence of SEQ ID No: 17, and an antibody having the _VL sequence of SEQ ID No: 22 and the _VH sequence of SEQ ID No: 27, respectively.

在一个技术方案中，本发明提供含有可变的V_L CDR1的CD38BP，该V_L CDR1实质上由与如SEQ ID No：3或SEQ ID No：13或SEQ IDNo：23所述的序列具有约50％，譬如至少60％、例如至少约70％、譬如至少约75％、例如至少约80％、譬如至少约85％、例如至少约90％、譬如至少约95％氨基酸序列同一性的序列所组成，其中CD38BP至少具有抗体抗原决定簇结合特征的实质部分(至少约50％、60％、70％、80％、90％、95％或更多)，该抗体分别具有SEQID No：3或SEQ ID No：13或SEQ ID No：23的可变V_L CDR1序列，譬如分别具有SEQ ID No：2或SEQ ID No：12或SEQ ID No：22的抗体，譬如分别具有SEQ ID No：2的V_L序列和SEQ IDNo：7的V_H序列的抗体，及分别具有SEQ ID No：12的V_L序列和SEQ ID No：17的V_H序列的抗体，和分别具有SEQ ID No：22的V_L序列和SEQ ID No：27的V_H序列的抗体。In one technical solution, the present invention provides CD38BP containing variable V _L CDR1, which V _L CDR1 is essentially composed of a sequence as described in SEQ ID No: 3 or SEQ ID No: 13 or SEQ ID No: 23 having about 50%, such as at least 60%, such as at least about 70%, such as at least about 75%, such as at least about 80%, such as at least about 85%, such as at least about 90%, such as at least about 95% amino acid sequence identity. Composition, wherein CD38BP has at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the epitope binding characteristics of an antibody having SEQ ID No: 3 or SEQ ID NO: The variable V _L CDR1 sequence of ID No: 13 or SEQ ID No: 23, such as an antibody having SEQ ID No: 2 or SEQ ID No: 12 or SEQ ID No: 22, respectively, such as having SEQ ID No: 2 respectively V _L sequence and the antibody of the V _H sequence of SEQ ID No: 7, and respectively have the antibody of the V _L sequence of SEQ ID No: 12 and the V _H sequence of SEQ ID No: 17, and respectively have the V of SEQ ID No: 22 Antibodies to the _L sequence and the _VH sequence of SEQ ID No: 27.

在一个技术方案中，本发明提供含有可变的V_L CDR2的CD38BP，该V_L CDR2实质上由与如SEQ ID No：4或SEQ ID No：14或SEQ IDNo：24所述的序列具有约50％，譬如至少60％、例如至少约70％、譬如至少约75％、例如至少约80％、譬如至少约85％、例如至少约90％、譬如至少约95％氨基酸序列同一性的序列所组成，其中CD38BP至少具有抗体抗原决定簇结合特征的实质部分(至少约50％、60％、70％、80％、90％、95％或更多)，该抗体分别具有SEQID No：4或SEQ ID No：14或SEQ ID No：24的可变V_L CDR2序列，譬如分别具有SEQ ID No：2或SEQ ID No：12或SEQ ID No：22的抗体，譬如分别具有SEQ ID No：2的V_L序列和SEQ IDNo：7的V_H序列的抗体，及分别具有SEQ ID No：12的V_L序列和SEQ ID No：17的V_H序列的抗体，和分别具有SEQ IDNo：22的V_L序列和SEQ ID No：27的V_H序列的抗体。In one technical solution, the present invention provides _CD38BP containing a variable V _L CDR2, which essentially consists of a sequence as described in SEQ ID No: 4 or SEQ ID No: 14 or SEQ ID No: 24 having about 50%, such as at least 60%, such as at least about 70%, such as at least about 75%, such as at least about 80%, such as at least about 85%, such as at least about 90%, such as at least about 95% amino acid sequence identity. Composition, wherein CD38BP has at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the epitope binding characteristics of an antibody having SEQ ID No: 4 or SEQ ID NO: 4 or SEQ ID NO: The variable V _L CDR2 sequence of ID No: 14 or SEQ ID No: 24, such as an antibody having SEQ ID No: 2 or SEQ ID No: 12 or SEQ ID No: 22, respectively, such as having SEQ ID No: 2 respectively Antibodies with _VL sequence and _VH sequence of SEQ ID No: 7, and antibodies with _VL sequence of SEQ ID No: 12 and _VH sequence of SEQ ID No: 17, respectively, and _VL with SEQ ID No: 22 sequence and the antibody of the _VH sequence of SEQ ID No: 27.

在一个技术方案中，本发明提供含有可变的V_L CDR3的CD38BP，该V_L CDR3实质上由与如SEQ ID No：5或SEQ ID No：15或SEQ IDNo：25所述的序列具有约50％，譬如至少60％、例如至少约70％、譬如至少约75％、例如至少约80％、譬如至少约85％、例如至少约90％、譬如至少约95％氨基酸序列同一性的序列所组成，其中CD38BP至少具有抗体抗原决定簇结合特征的实质部分(至少约50％、60％、70％、80％、90％、95％或更多)，该抗体分别具有SEQID No：5或SEQ ID No：15或SEQ ID No：25的可变V_L CDR3序列，譬如分别具有SEQ ID No：2或SEQ ID No：12或SEQ ID No：22的抗体，譬如分别具有SEQ ID No：2的V_L序列和SEQ IDNo：7的V_H序列的抗体，及分别具有SEQ ID No：12的V_L序列和SEQ ID No：17的V_H序列的抗体，和分别具有SEQ IDNo：22的V_L序列和SEQ ID No：27的V_H序列的抗体。In one technical solution, the present invention provides _CD38BP containing variable V _L CDR3, which essentially consists of a sequence as described in SEQ ID No: 5 or SEQ ID No: 15 or SEQ ID No: 25 having about 50%, such as at least 60%, such as at least about 70%, such as at least about 75%, such as at least about 80%, such as at least about 85%, such as at least about 90%, such as at least about 95% amino acid sequence identity. Composition, wherein CD38BP has at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the epitope binding characteristics of an antibody having SEQ ID No: 5 or SEQ ID No: 5 or SEQ ID NO: The variable V _L CDR3 sequence of ID No: 15 or SEQ ID No: 25, such as an antibody having SEQ ID No: 2 or SEQ ID No: 12 or SEQ ID No: 22, respectively, such as having SEQ ID No: 2 respectively Antibodies with _VL sequence and _VH sequence of SEQ ID No: 7, and antibodies with _VL sequence of SEQ ID No: 12 and _VH sequence of SEQ ID No: 17, respectively, and _VL with SEQ ID No: 22 sequence and the antibody of the _VH sequence of SEQ ID No: 27.

在一个技术方案中，本发明提供含有可变的V_H的CD38BP，该V_H 实质上由与如SEQID No：7或SEQ ID No：17或SEQ ID No：27所述的序列具有约50％，譬如至少60％、例如至少约70％、譬如至少约75％、例如至少约80％、譬如至少约85％、例如至少约90％、譬如至少约95％氨基酸序列同一性的序列所组成，其中CD38BP至少具有抗体抗原决定簇结合特征的实质部分(至少约50％、60％、70％、80％、90％、95％或更多)，该抗体分别具有SEQ ID No：7或SEQ ID No：17或SEQ ID No： 27的可变V_L CDR3序列，譬如分别具有SEQ ID No：7的V_H序列和SEQID No：2的V_L序列的抗体，及分别具有SEQ ID No：17的V_H序列和SEQ ID No：12的V_L序列的抗体，和分别具有SEQ ID No：27的V_H序列和SEQ ID No：22的V_L序列的抗体。In one technical solution, the present invention provides _CD38BP containing a variable V _H , which is substantially composed of about 50% of the sequence described in SEQ ID No: 7 or SEQ ID No: 17 or SEQ ID No: 27 , such as at least 60%, such as at least about 70%, such as at least about 75%, such as at least about 80%, such as at least about 85%, such as at least about 90%, such as at least about 95% amino acid sequence identity. wherein the CD38BP has at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the epitope binding characteristics of an antibody having SEQ ID No: 7 or SEQ ID No: 17 or the variable V _L CDR3 sequence of SEQ ID No: 27, such as an antibody having a V _H sequence of SEQ ID No: 7 and a V L sequence of SEQ ID No: 2, respectively, and an antibody having a V _L sequence of SEQ ID No: 17, respectively An antibody having a _VH sequence and a _VL sequence of SEQ ID No: 12, and an antibody having a _VH sequence of SEQ ID No: 27 and a _VL sequence of SEQ ID No: 22, respectively.

在一个技术方案中，本发明提供含有可变的V_H CDR1的CD38BP，该V_H CDR1实质上由与如SEQ ID No：8或SEQ ID No：18或SEQ IDNo：28所述的序列具有约50％，譬如至少60％、例如至少约70％、譬如至少约75％、例如至少约80％、譬如至少约85％、例如至少约90％、譬如至少约95％氨基酸序列同一性的序列所组成，其中CD38BP至少具有抗体抗原决定簇结合特征的实质部分(至少约50％、60％、70％、80％、90％、95％或更多)，该抗体分别具有SEQID No：8或SEQ ID No：18或SEQ ID No：28的可变V_H CDR1序列，譬如分别具有SEQ ID No：7或SEQ ID No：17或SEQ ID No：27的抗体，譬如分别具有SEQ ID No：7的V_H序列和SEQ IDNo：2的V_L序列的抗体，及分别具有SEQ ID No：17的V_H序列和SEQ ID No：12的V_L序列的抗体，和分别具有SEQ IDNo：27的V_H序列和SEQ ID No：22的V_L序列的抗体。In one technical solution, the present invention provides CD38BP containing variable _VH CDR1, which _VH CDR1 is essentially composed of a sequence as described in SEQ ID No: 8 or SEQ ID No: 18 or SEQ ID No: 28 having about 50%, such as at least 60%, such as at least about 70%, such as at least about 75%, such as at least about 80%, such as at least about 85%, such as at least about 90%, such as at least about 95% amino acid sequence identity. Composition, wherein CD38BP has at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the epitope binding characteristics of an antibody having SEQ ID No: 8 or SEQ ID No: 8 or SEQ ID NO: The variable _VH CDR1 sequence of ID No: 18 or SEQ ID No: 28, such as an antibody having SEQ ID No: 7 or SEQ ID No: 17 or SEQ ID No: 27, respectively, such as having SEQ ID No: 7, respectively Antibodies with _VH sequence and _VL sequence of SEQ ID No: 2, and antibodies with _VH sequence of SEQ ID No: 17 and _VL sequence of SEQ ID No: 12, respectively, and _VH with SEQ ID No: 27 sequence and the antibody to the _VL sequence of SEQ ID No: 22.

在一个技术方案中，本发明提供含有可变的V_H CDR2的CD38BP，该V_H CDR2实质上由与如SEQ ID No：9或SEQ ID No：19或SEQ IDNo：29所述的序列具有约50％，譬如至少60％、例如至少约70％、譬如至少约75％、例如至少约80％、譬如至少约85％、例如至少约90％、譬如至少约95％氨基酸序列同一性的序列所组成，其中CD38BP至少具有抗体抗原决定簇结合特征的实质部分(至少约50％、60％、70％、80％、90％、95％或更多)，该抗体分别具有SEQID No：9或SEQ ID No：19或SEQ ID No：29的可变V_H CDR2序列，譬如分别具有SEQ ID No：7或SEQ ID No：17或SEQ ID No：27的抗体，譬如分别具有SEQ ID No：7的V_H序列和SEQ IDNo：2的V_L序列的抗体，及分别具有SEQ ID No：17的V_H序列和SEQ ID No：12的V_L序列的抗体，和分别具有SEQ IDNo：27的V_H序列和SEQ ID No：22的V_L序列的抗体。In one technical scheme, the present invention provides CD38BP containing variable _VH CDR2, which _VH CDR2 is essentially composed of a sequence as described in SEQ ID No: 9 or SEQ ID No: 19 or SEQ ID No: 29 having about 50%, such as at least 60%, such as at least about 70%, such as at least about 75%, such as at least about 80%, such as at least about 85%, such as at least about 90%, such as at least about 95% amino acid sequence identity. Composition, wherein CD38BP has at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the antigenic determinant binding characteristics of an antibody having SEQ ID No: 9 or SEQ ID No: 9 or SEQ ID NO: The variable _VH CDR2 sequence of ID No: 19 or SEQ ID No: 29, such as an antibody having SEQ ID No: 7 or SEQ ID No: 17 or SEQ ID No: 27, respectively, such as having SEQ ID No: 7, respectively Antibodies with _VH sequence and _VL sequence of SEQ ID No: 2, and antibodies with _VH sequence of SEQ ID No: 17 and _VL sequence of SEQ ID No: 12, respectively, and _VH with SEQ ID No: 27 sequence and the antibody to the _VL sequence of SEQ ID No: 22.

在一个技术方案中，本发明提供含有可变的V_H CDR3的CD38BP，该V_H CDR3实质上由与如SEQ ID No：10或SEQ ID No：20或SEQ IDNo：30所述的序列具有约50％，譬如至少60％、例如至少约70％、譬如至少约75％、例如至少约80％、譬如至少约85％、例如至少约90％、譬如至少约95％氨基酸序列同一性的序列所组成，其中CD38BP至少具有抗体抗原决定簇结合特征的实质部分(至少约50％、60％、70％、80％、90％、95％或更多)，该抗体分别具有SEQID No：10或SEQ ID No：20或SEQ ID No：30的可变V_H CDR3序列，譬如分别具有SEQ ID No：7或SEQ ID No：17或SEQ ID No：27的抗体，譬如分别具有SEQ ID No：7的V_H序列和SEQ IDNo：2的V_L序列的抗体，及分别具有SEQ ID No：17的V_H序列和SEQ ID No：12的V_L序列的抗体，和分别具有SEQ IDNo：27的V_H序列和SEQ ID No：22的V_L序列的抗体。In one technical solution, the present invention provides CD38BP containing variable _VH CDR3, which _VH CDR3 is essentially composed of a sequence as described in SEQ ID No: 10 or SEQ ID No: 20 or SEQ ID No: 30 having about 50%, such as at least 60%, such as at least about 70%, such as at least about 75%, such as at least about 80%, such as at least about 85%, such as at least about 90%, such as at least about 95% amino acid sequence identity. Composition, wherein CD38BP has at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the antigenic determinant binding characteristics of an antibody having SEQ ID No: 10 or SEQ ID No: 10 or SEQ ID NO: The variable _VH CDR3 sequence of ID No: 20 or SEQ ID No: 30, such as an antibody having SEQ ID No: 7 or SEQ ID No: 17 or SEQ ID No: 27, respectively, such as having SEQ ID No: 7 respectively Antibodies with _VH sequence and _VL sequence of SEQ ID No: 2, and antibodies with _VH sequence of SEQ ID No: 17 and _VL sequence of SEQ ID No: 12, respectively, and _VH with SEQ ID No: 27 sequence and the antibody to the _VL sequence of SEQ ID No: 22.

两个序列间的百分比同一性是序列中相同位点的数目的函数(即，％同源性＝#一致位点/总#位点×100)，为了对两个序列进行最适比对，需要考虑空位数目和每个空位的长度。可使用数学算法来实现两个序列间的序列比较和百分比同一性的测定，如下非限制性实施例所述。The percent identity between two sequences is a function of the number of identical positions in the sequences (i.e., % Homology = #identical positions/total # positions x 100), for optimal alignment of two sequences, The number of slots and the length of each slot need to be considered. The comparison of sequences and the determination of percent identity between two sequences can be accomplished using a mathematical algorithm, as described in the following non-limiting examples.

两个核苷酸序列间的百分比同一性可使用GCG软件包(可从 http://www.gcg.com获得)中的GAP程序，通过NWSgapdha.CMP矩阵和40、50、70或80的空位权重及1、2、3、4、5或6的长度权重来测定。两个核苷酸或氨基酸序列间的百分比同一性也可使用E.MeyersandW.Miller，Comput.Appl.Biosci 4，11-17(1988))中的算法来测定，它已整合到ALIGN程序(2.0版)，所用的是PAM120权重残基表，空位长度罚分为12，空位罚分为4。另外，两个氨基酸序列间的百分比同一性可使用Needleman and Wunsch，J.MoI.Biol.48，444-453(1970))算法来测定，它已整合到GCG软件包(可从http://www.gcg.com获得)中的GAP程序中，使用的是Blossum 62矩阵和PAM250矩阵，空位权重为16、14、12、10、8、6或4，长度权重为1、2、3、4、5或6。The percent identity between two nucleotide sequences can be determined using the GAP program in the GCG software package (available from http://www.gcg.com ) by using the NWSgapdha.CMP matrix and gaps of 40, 50, 70 or 80 Weights and length weights of 1, 2, 3, 4, 5 or 6 are determined. The percent identity between two nucleotide or amino acid sequences can also be determined using the algorithm in E. Meyers and W. Miller, Comput. Appl. Biosci 4, 11-17 (1988)), which has been incorporated into the ALIGN program (2.0 version), the PAM120 weighted residue table was used, the gap length penalty was 12, and the gap penalty was 4. Alternatively, the percent identity between two amino acid sequences can be determined using the Needleman and Wunsch, J. MoI. Biol. 48, 444-453 (1970)) algorithm, which has been incorporated into the GCG software package (available from http:// In the GAP program obtained from www.gcg.com ), Blossum 62 matrix and PAM250 matrix are used, the gap weight is 16, 14, 12, 10, 8, 6 or 4, and the length weight is 1, 2, 3, 4 , 5 or 6.

本发明的核酸和蛋白序列可进一步用作“查询序列”，以对公共数据库进行搜索，例如鉴定相关序列。这种搜索可使用Altschul et al.，J.MoI.Biol.215，403-10(1990)NBLAST和XBLAST程序(2.0版)来执行。可使用NBLAST程序，分数＝100，字长＝12来执行BLAST核苷酸搜索，以获得与本发明中核酸分子同源的核苷酸序列。可使用XBLAST程序，分数＝50，字长＝3来执行BLAST蛋白搜索，以获得与本发明中蛋白分子同源的氨基酸序列。为了获得空位比对以进行比较，可使用如Altschulet al.，Nucleic Acids Res.25(17)，3389-3402(1997)所述的GappedBLAST。当使用BLAST和Gapped BLAST程序时，可使用各自程序(例 如XBLAST和NBLAST)的缺省参数。参见http://www.ncbi.nlm.nih.gov。The nucleic acid and protein sequences of the invention can further be used as "query sequences" to perform searches against public databases, eg, to identify related sequences. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul et al., J. MoI. Biol. 215, 403-10 (1990). BLAST nucleotide searches can be performed using the NBLAST program, score=100, wordlength=12 to obtain nucleotide sequences homologous to nucleic acid molecules of the invention. BLAST protein searches can be performed with the XBLAST program, score = 50, wordlength = 3 to obtain amino acid sequences homologous to protein molecules of the invention. To obtain gapped alignments for comparison, GappedBLAST as described by Altschule et al., Nucleic Acids Res. 25(17), 3389-3402 (1997) can be used. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (eg, XBLAST and NBLAST) can be used. See http://www.ncbi.nlm.nih.gov .

CDR变异体的序列可通过大部分保守替代而与母抗体序列的CDR序列有所不同；例如，变异体中至少约35％、约50％或更多、约60％或更多、约70％或更多、约75％或更多、约80％或更多、约85％或更多、约90％或更多、约95％或更多(譬如约65-99％)的替代是保守氨基酸残基的替换。在本发明的语境中，保守替代定义为如下3个表的一个或多个中所表示的氨基酸种类中的替代：The sequence of the CDR variant may differ from that of the parent antibody sequence by mostly conservative substitutions; for example, at least about 35%, about 50% or more, about 60% or more, about 70% of the variant or more, about 75% or more, about 80% or more, about 85% or more, about 90% or more, about 95% or more (such as about 65-99%) of the substitutions are conservative Amino acid residue substitutions. In the context of the present invention, conservative substitutions are defined as substitutions in the amino acid classes indicated in one or more of the following 3 tables:

保守替代的氨基酸残基种类Conservatively substituted amino acid residues

可选择的保守氨基酸残基替代的种类Types of alternative conservative amino acid residue substitutions

更保守的替代组包括：缬氨酸-亮氨酸-异亮氨酸、苯丙氨酸-酪氨酸、赖氨酸-精氨酸、丙氨酸-缬氨酸和天冬氨酸-谷氨酸。其它组氨基酸也可通过如Creighton(1984)Proteins：Structure and Molecular Properties(2d Ed.1993)，W.H.Freeman andCompany所述的原理来表示。More conservative substitution groups include: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, and aspartate- glutamic acid. Other groups of amino acids can also be represented by principles such as those described by Creighton (1984) Proteins: Structure and Molecular Properties (2d Ed. 1993), W.H. Freeman and Company.

在本发明的一个技术方案中，与实施例中抗体的CDR相比较，在变异的CDR中，根据亲水/疏水性质和残基分子量/大小的保守性实质上也是保留的(例如，序列的分子量种类、亲水得分或二者保留了至少约50％、至少约60％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％、至少约95％或更多(例如65-99％))。例如，保守残基的替代也可选择性地基于根据保守基团分子量的强或弱进行的替代，这在本领域是已知的。In one technical solution of the present invention, compared with the CDR of the antibody in the examples, in the variant CDR, the conservation according to the hydrophilic/hydrophobic properties and the molecular weight/size of the residues is also substantially preserved (for example, sequence At least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95% of the molecular weight species, hydrophilic score, or both are retained or more (eg 65-99%)). For example, substitution of conserved residues may also optionally be based on substitution based on the strength or weakness of the molecular weight of the conserved group, as is known in the art.

选择性地，也可通过相似性得分，例如通过使用BLAST程序(例如，可通过NCBI获得BLAST 2.2.8)所确定的那样来测定相似残基的 保持能力。典型地，合适的变异体与母肽具有至少约45％，譬如至少约55％、至少约65％、至少约75％、至少约85％、至少约90％、至少约95％或更高(例如约70-99％)的相似性。Alternatively, the retention of similar residues can also be determined by a similarity score, e.g., as determined using the BLAST program (e.g., BLAST 2.2.8 available through NCBI). Typically, suitable variants have at least about 45%, such as at least about 55%, at least about 65%, at least about 75%, at least about 85%, at least about 90%, at least about 95% or higher ( eg about 70-99%) similarity.

可通过选择与上述已定义的组中具有更低保守性的替代来产生功能上的实质改变。例如，可产生在改变的区域更显著地影响肽的结构的非保守性替代，例如，α-螺旋或β-折叠；在靶位点上分子的电荷或疏水性或侧链的位阻。通常期望在多肽的性质上产生最大改变的替代是那些，1)亲水残基，例如丝氨酰或苏氨酰替换为(或被替换)疏水残基，例如亮氨酰、异亮氨酰、苯丙氨酰、缬氨酰或丙氨酰；2)半胱氨酸或脯氨酸替换为(或被替换)任何其它残基；3)具有正电侧链的残基，例如赖氨酰、精氨酰或组氨酰替换为(或被替换)负电残基，例如谷氨酰或天冬氨酰；或4)具有空间位阻侧链的残基，例如苯丙氨酸替换为(或被替换)没有侧链的残基，例如甘氨酸。因此，这些及其它非保守替代可引入到其中功能/结构需要显著改变的肽变异体中，而且这种消除了结构/功能的保守性的改变是所期望的。Substantial changes in function can be made by selecting less conservative substitutions from the above defined groups. For example, non-conservative substitutions can be made that more significantly affect the structure of the peptide in altered regions, eg, α-helices or β-sheets; the charge or hydrophobicity of the molecule or steric hindrance of side chains at the target site. The substitutions that are generally expected to produce the greatest change in the properties of the polypeptide are those, 1) a hydrophilic residue such as seryl or threonyl is replaced (or replaced) by a hydrophobic residue such as leucyl, isoleucyl , phenylalanyl, valyl, or alanyl; 2) cysteine or proline is replaced (or replaced) by any other residue; 3) residues with positively charged side chains, such as lysine acyl, arginyl or histidyl are replaced (or replaced) by negatively charged residues such as glutamyl or aspartyl; or 4) residues with sterically hindered side chains such as phenylalanine are replaced by (or to be substituted for) a residue without a side chain, such as glycine. Thus, these and other non-conservative substitutions can be introduced into peptide variants where a significant change in function/structure is desired, and such changes that eliminate the conservative nature of structure/function are desired.

产生替代变异体的简便途径是利用本领域中已知方法使用噬菌体而进行的亲和力成熟技术。为了鉴定候选的高变区域位点以进行修饰，可进行丙氨酸扫描突变方法，以鉴定对抗原结合具有显著贡献的高变区域。选择性或附加地，分析抗原-抗体复合物的晶体结构以鉴定抗原-抗体间的接触位点是有益的。这些接触的残基和临近的残基可能是替代的合适候选者。A convenient way to generate substitutional variants is affinity maturation using phage by methods known in the art. To identify candidate hypervariable region sites for modification, an alanine scanning mutagenesis approach can be performed to identify hypervariable regions that contribute significantly to antigen binding. Alternatively or additionally, it may be beneficial to analyze the crystal structure of the antigen-antibody complex to identify antigen-antibody contact sites. These contact residues and neighboring residues may be suitable candidates for substitution.

当通过高变区插入以生成变异抗体时，应该考虑已知抗体中所研究的高变区长度的典型范围。例如，对于轻链可变结构域的第一个高变区而言，可在保持实质相似性及所预期的合适大小的情况下，向母抗体V_L CDR1中引入插入，根据Kabat等，同上，典型地V_L CDR1具有总共约9-20(例如，约10-17)个残基。同样，典型地V_L CDR2具有总共长度约5-10个残基；典型地V_L CDR3有约7-20个残基长；典型地V_H CDR1有约10-15个残基长；典型地V_H CDR2有约15-20个残基长；典型地V_H CDR3有约6-30个残基长(例如，3-25残基)。使用Kabat中所述的比对和计数方法，典型地，在V_H CDR3中生成位于V_H区域中的插入，而且典型地，该插入靠近结构域的C末端，譬如，在母V_H CDR3的约残基97-102处(例如毗邻，或C末端邻接到，母V_HCDR3序列的100号 残基)。可随机制备在其高变区有插入氨基酸残基的抗体变异体，特别地，其中母抗体针对靶抗原的起始结合亲和力可使随机产生的抗体变异体易于被筛选。例如，噬菌体展示提供了一种筛选这种随机变异体的方法。When generating variant antibodies through hypervariable region insertions, the typical range of hypervariable region lengths studied in known antibodies should be considered. For example, for the first hypervariable region of the light chain variable domain, an insertion can be introduced into the _VL CDR1 of the parent antibody while maintaining substantial similarity and expected appropriate size, according to Kabat et al., supra Typically, a V _L CDR1 has a total of about 9-20 (eg, about 10-17) residues. Likewise, typically _VL CDR2s have a total length of about 5-10 residues; typically _VL CDR3s are about 7-20 residues long; typically _VH CDR1s are about 10-15 residues long; typically _VH CDR2s are about 15-20 residues long; typically _VH CDR3s are about 6-30 residues long (eg, 3-25 residues). Using the alignment and counting methods described in Kabat, an insertion is typically generated in a _VH CDR3 that is located in the _VH region, and typically this insertion is near the C-terminus of the domain, for example, at the end of the parent _VH CDR3. At about residues 97-102 (eg, adjacent to, or C-terminally adjacent to, residue 100 of the parent _VH CDR3 sequence). Antibody variants having inserted amino acid residues in their hypervariable regions can be generated randomly, particularly where the initial binding affinity of the parent antibody for the target antigen allows the randomly generated antibody variants to be readily screened. For example, phage display provides a method to screen for such random variants.

在设计、构建和/或评估CDR变异体时，应注意到可改变CDR区域，使其与抗原决定簇更好结合。典型地，可通过提供互补表面，可能包括可伸出到抗原蛋白表面的指状结构或其它固定抗原决定簇的抗体决定簇结构来操作抗体CDRs。如果抗原决定簇没有紧密固定，抗体将不会发挥最大亲和力。但对于抗原决定簇而言，在抗体决定簇结构中，通常是几个关键残基来主要负责这种结合。因此，CDR序列在针对相同肽的抗体间，可在长度和组成上有显著差异。技术人员知道诸如酪氨酸残基(例如，位于V_H CDR3序列中)这样的通常对于这种抗原决定簇结合具有显著贡献的特定残基典型地保留在CDR变异体中。When designing, constructing, and/or evaluating CDR variants, it should be noted that CDR regions can be altered to better bind epitopes. Typically, antibody CDRs can be manipulated by providing complementary surfaces, possibly including finger-like structures that can protrude to the surface of the antigenic protein, or other antibody determinant structures that immobilize the antigenic determinant. If the epitope is not tightly anchored, the antibody will not exert maximum affinity. But for antigenic determinants, in the antibody determinant structure, usually a few key residues are mainly responsible for this binding. Thus, CDR sequences can vary significantly in length and composition between antibodies raised against the same peptide. The skilled person knows that certain residues, such as tyrosine residues (eg, located in the _VH CDR3 sequence), which normally contribute significantly to such antigenic determinant binding, are typically retained in CDR variants.

与抗原和母抗体间的氨基酸接触相比较，通过引入一个或多个增加抗原中存在的一个或多个氨基酸残基与抗体中存在的一个或多个氨基酸残基间的接触或在能量方面有利的相互作用的氨基酸残基(通过替代或插入)，则CDR区域的变异体也可增加抗原与抗体变异体间的氨基酸接触。目的氨基酸相互作用可从疏水键相互作用、范德华相互作用和离子相互作用中选择。Increased or energetically favored contact between one or more amino acid residues present in the antigen and one or more amino acid residues present in the antibody by introducing one or more Variants in the CDR regions can also increase the amino acid contacts between the antigen and antibody variants. The amino acid interaction of interest can be selected from hydrophobic bond interaction, van der Waals interaction, and ionic interaction.

本领域技术人员要注意在设计和筛选含本发明抗体的CDR变异体的CD38BP时的其它原则。Those skilled in the art are aware of additional principles in designing and screening CD38BPs containing CDR variants of the antibodies of the invention.

对于作为实施例中的抗体的CDRs变异体的CDR变异体而言，特别是对抗CD38抗体或其片段中的变异CDR而言，典型地，应保留支持和/或定向CDR结构性环结构所需的残基；典型地，也不改变位于CDR结构性环约10埃(但选择性地只包括该区域具有约5平方埃或更大的水溶剂接触表面的残基)的残基或仅通过保守氨基酸残基替代进行改变；和/或典型地，氨基酸序列仅具有有限数量的插入和/或缺失(即便要)，从而使CDR结构性类似环的结构可在变异体中保留(相关技术和相关原理的描述参见，例如Schiweck et al.，J MoIBiol.268(5)，934-51(1997)、Morea，Biophys Chem.68(1-3)，9-16(1997)、Shirai et al.，FEBS Lett.399(1-2)，1-8(1996)、Shirai et al.，FEBS Lett.455(1-2)，188-97(1999)、Reckzo et al.，Protein Eng.8(4)，389-95(1995)and Eigenbrot et al.，J MoIBiol.229(4)，969-95(1993))。也参见WO03/048185、WO 03/070747和WO 03/027246。For CDR variants that are variants of the CDRs of the antibodies of the examples, particularly variant CDRs in anti-CD38 antibodies or fragments thereof, typically the structural loop structures required to support and/or orient the CDRs should be retained. residues; typically, residues located within about 10 angstroms of the CDR structural loops (but selectively only including residues with about 5 square angstroms or greater of water-solvent contacting surface in this region) are also not altered or only by and/or typically, the amino acid sequence has only a limited number of insertions and/or deletions, if any, so that the CDRs are structurally loop-like structures retained in variants (related art and For a description of the relevant principles see, for example, Schiweck et al., J MoI Biol. 268(5), 934-51 (1997), Morea, Biophys Chem. 68(1-3), 9-16 (1997), Shirai et al. , FEBS Lett.399(1-2), 1-8(1996), Shirai et al., FEBS Lett.455(1-2), 188-97(1999), Reckzo et al., Protein Eng.8( 4), 389-95(1995) and Eigenbrot et al., J MoI Biol. 229(4), 969-95(1993)). See also WO 03/048185, WO 03/070747 and WO 03/027246.

其它技术也可用于产生变异的抗体，包括定向进化和其它变异体产生技术，例如在US 20040009498，Marks et al.，Methods MoI Biol.248，327-43(2004)、Azriei-Rosenfeld et al.，J MoI Biol.335(1)，177-92(2004)、Park et al.，Biochem BiophysRes Commun.275(2).553-7(2000)、Kang et al.，Proc Natl Acad Sci USA.88(24)，11120-3(1991)、Zahnd et al.，J Biol Chem.279(18)，18870-7(2004)、Xu et al.，ChemBiol.9(8)，933-42(2002)、Border et al.，Proc Natl Acad Sci USA.97(20)，10701-5(2000)、Crameri et al.，Nat Med.2(1)，100-2(1996)中所述，更普遍地，例如在WO 03/048185中所述。Other techniques can also be used to generate variant antibodies, including directed evolution and other variant generation techniques, for example in US 20040009498, Marks et al., Methods MoI Biol. 248, 327-43 (2004), Azriei-Rosenfeld et al., J MoI Biol.335(1), 177-92(2004), Park et al., Biochem BiophysRes Commun.275(2).553-7(2000), Kang et al., Proc Natl Acad Sci USA.88( 24), 11120-3(1991), Zahnd et al., J Biol Chem.279(18), 18870-7(2004), Xu et al., ChemBiol.9(8), 933-42(2002), Border et al., Proc Natl Acad Sci USA. 97(20), 10701-5 (2000), Crameri et al., Nat Med. 2(1), 100-2 (1996), and more generally, For example as described in WO 03/048185.

产生的抗体变异体可用于任何合适的筛选技术，而且可筛选在一个或多个相关检测中具有合适的和所需的高级特征的抗体，用于进一步研究。The antibody variants generated can be used in any suitable screening technique, and antibodies with suitable and desired high-level characteristics in one or more relevant assays can be screened for further study.

如上所述的含CDR序列的CD38BPs在保留至少实质部分(至少约50％、60％、70％、80％、90％、95％或更多)具有如SEQ ID No：2中的V_L序列和如SEQ ID No：7中的V_H序列的抗体，和/或具有如SEQ IDNo：12中的V_L序列和如SEQ ID No：17中的V_H序列的抗体，和/或具有如SEQ ID No：22中的V_L序列和如SEQ ID No：27中的V_H序列的抗体的亲和性/强度和/或特异性/选择性的情况下，可含有任何合适数目和组合的这种V_L和V_H CDRs，但可选择性地，在诸如人患者中的免疫原性、对抗原决定簇的亲和性、增加的半衰期等这样的其它特性上会有所差别。在某些情况下，这种CD38BP可比母抗体具有更高的亲和性、选择性和/或特异性。在一个技术方案中，在本发明的CD38BP中可存在少于1个完整系列的V_L CDRs和/或V_H CDRs。在一个技术方案中，存在所有的V_L CDRs和V_H CDRs。The above-mentioned CD38BPs containing CDR sequence have the V _L sequence in SEQ ID No: 2 while retaining at least a substantial part (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) And as the antibody of the _VH sequence among the SEQ ID No:7, and/or have as the antibody of the _VL sequence among the SEQ IDNo:12 and as the _VH sequence among the SEQ ID No:17, and/or have as SEQ ID No: In the case of the affinity/strength and/or specificity/selectivity of the _VL sequence among ID No:22 and the antibody such as the _VH sequence among SEQ ID No:27, any suitable number and combination of these may be contained. The _VL and _VH CDRs, but optionally differ in other properties such as immunogenicity, affinity for antigenic determinants, increased half-life, etc. in human patients. In certain instances, such CD38BP may have higher affinity, selectivity and/or specificity than the parent antibody. In one embodiment, there may be less than 1 complete set of V _L CDRs and/or V _H CDRs in the CD38BP of the present invention. In one embodiment, all _VL CDRs and _VH CDRs are present.

与-003和-005及-024相比较，在本发明的变异CD38BPs中可改变或保留的抗体的其它功能性质的实施例有：Compared with -003 and -005 and -024, examples of other functional properties of antibodies that can be changed or retained in the variant CD38BPs of the present invention are:

(1)与CD38结合的高亲和性；(1) High affinity binding to CD38;

(2)从CD38的低解离速率；(2) low dissociation rate from CD38;

(3)抑制或阻断CD38与CD38靶的结合；(3) Inhibit or block the binding of CD38 to CD38 target;

(4)清除表达CD38的T细胞或B细胞；(4) Eliminate T cells or B cells expressing CD38;

(5)诱导高水平的CD55/59阴性或CD55/59阳性细胞的CDC；(5) CDCs that induce high levels of CD55/59 negative or CD55/59 positive cells;

(6)与CD38结合后转位到脂质筏中；(6) Translocate into lipid rafts after combining with CD38;

(7)T细胞耐受性；(7) T cell tolerance;

(8)抑制表达CD38的T或B细胞的增殖；(8) Inhibit the proliferation of T or B cells expressing CD38;

(9)CD38内在化；(9) CD38 internalization;

(10)抑制或诱导CD38的酶活性；(10) Inhibit or induce the enzymatic activity of CD38;

(11)抑制或诱导CD38诱导的信号转导；(11) Inhibit or induce CD38-induced signal transduction;

(12)诱导或抑制细胞因子的产生；(12) Inducing or inhibiting the production of cytokines;

(13)诱导或阻断T细胞或B细胞的分化；(13) Inducing or blocking the differentiation of T cells or B cells;

(14)诱导或从凋亡中恢复；(14) induction or recovery from apoptosis;

(15)减轻或增加NK细胞的裂解诱导性；(15) Reduce or increase the lysis inducibility of NK cells;

(16)诱导或抑制胰腺中β细胞中胰岛素的产生；(16) Inducing or inhibiting the production of insulin in β cells in the pancreas;

(17)延长具有表达CD38的肿瘤细胞的患者的存活；和/或(17) prolong the survival of patients with tumor cells expressing CD38; and/or

(18)当与适量的效应细胞混合时诱导CD38靶的ADCC。本发明也提供CD38BPs，其特征在于它们可与具有SEQ ID No：2的V_L序列和如SEQ ID No：7中的V_H序列的抗体(譬如抗体-003)，或具有如SEQID No：12中的V_L序列和如SEQ ID No：17中的V_H序列的抗体(譬如抗体-005)，或具有如SEQ ID No：22中的V_L序列和如SEQ ID No：27中的V_H序列的抗体(譬如抗体-024)竞争(竞争性抑制)或交叉竞争(即，相对部分抑制抗原决定簇的结合)结合CD38。(18) Induces ADCC of CD38 targets when mixed with appropriate amount of effector cells. The present invention also provides CD38BPs, characterized in that they can be combined with an antibody (such as Antibody-003) having a V _L sequence of SEQ ID No: 2 and a V _H sequence such as SEQ ID No: 7, or an antibody having a V L sequence such as SEQ ID No: 12 An antibody (such as Antibody-005) having a _VL sequence as in SEQ ID No: 17 and a _VH sequence as in SEQ ID No: 17, or has a _VL sequence as in SEQ ID No: 22 and a _VH as in SEQ ID No: 27 Antibodies of the sequence (eg Antibody-024) compete (competitive inhibition) or cross-compete (ie, relatively partially inhibit the binding of the epitope) for binding to CD38.

例如，这种CD38BP可以是来源于抗体的Fab片段，该抗体可结合到与具有如SEQ IDNo：2中的V_L序列和如SEQ ID No：7中的V_H序列的抗体，和/或具有如SEQ ID No：12中的V_L序列和如SEQ ID No：17中的V_H序列的抗体，和/或具有如SEQ ID No：22中的V_L序列和如SEQIDNo：27中的V_H序列的抗体相结合的抗原决定簇相同或重叠的抗原决定簇上。这种Fab片段由于其与mAb分子相比较具有较小的尺寸，因此不能显著性地与上述抗体竞争结合到CD38时，虽然其所来源的抗体可以做到这一点。然而，这种CD38BP在靠近CD38区域的类似定位中也有作用(例如，在免疫连接物的CD38BP中的细胞毒素、放射性核苷等的定位)。所以，这种CD38BPs可用于本发明的方法中，因此也提供在本发明中。For example, such CD38BP may be a Fab fragment derived from an antibody that binds to an antibody having a V _L sequence such as SEQ ID No: 2 and a V _H sequence such as SEQ ID No: 7, and/or has An antibody having a _VL sequence as in SEQ ID No: 12 and a _VH sequence as in SEQ ID No: 17, and/or has a _VL sequence as in SEQ ID No: 22 and a _VH as in SEQ ID No: 27 Sequences of antibodies that bind to the same epitope or on overlapping epitopes. Due to its small size compared to the mAb molecule, this Fab fragment cannot significantly compete with the above-mentioned antibody for binding to CD38, although the antibody from which it is derived can do so. However, this CD38BP also plays a role in similar localization near CD38 regions (eg, localization of cytotoxins, radionucleosides, etc. in CD38BP of immunolinkers). Therefore, such CD38BPs can be used in the methods of the present invention and are therefore also provided in the present invention.

可通过任何合适的技术来测定两个或多个CD38BPs竞争结合到 CD38或CD38的一部分。在一个技术方案中，例如，竞争作用如实施例7、8和9中所述进行测定。The competition of two or more CD38BPs for binding to CD38 or a portion of CD38 can be determined by any suitable technique. In one embodiment, competition is determined as described in Examples 7, 8 and 9, for example.

本发明语境中的竞争作用是指特定分子与特定结合伴侣之间的结合倾向性在存在另一种可结合到结合伴侣的分子时有任何可检测的显著降低。典型地，竞争是指CD38BP与以下物质的结合在存在另一种CD38BP时，使用足量的两种或多种竞争性CD38BPs和CD38BPs分子通过例如ELISA分析或FACS分析(如实施例部分所述)进行测定，至少约有10％的降低，譬如至少约15、或至少约20％的降低Competition in the context of the present invention refers to any detectable significant decrease in the binding propensity of a particular molecule to a particular binding partner in the presence of another molecule which can bind to the binding partner. Typically, competition refers to the binding of a CD38BP to, in the presence of another CD38BP, the use of sufficient quantities of two or more competing CD38BPs and CD38BPs molecules by, for example, ELISA analysis or FACS analysis (as described in the Examples section) A reduction of at least about 10%, such as a reduction of at least about 15, or at least about 20%, is determined

(a)CD38形式(例如，“加工过的”、“成熟的”、“未加工的”、“没加工的”或“不成熟的”CD38)；(a) CD38 form (eg, "processed", "mature", "unprocessed", "unprocessed" or "immature" CD38);

(b)游离CD38形式(例如，通过体内加工产生的CD38片段)；(b) Free CD38 forms (e.g., CD38 fragments produced by in vivo processing);

(c)由诸如CD31这样的与CD38相关的另一种肽和CD38组成的异源二肽；(c) a heterologous dipeptide consisting of another peptide related to CD38, such as CD31, and CD38;

(d)CD38和诸如cAMP、NAD+和/或cADPR这样的一种或多种底物组成的复合物；(d) a complex of CD38 and one or more substrates such as cAMP, NAD+ and/or cADPR;

(e)CD38与诸如CD31这样的可溶性配体组成的二聚体化的，相关联的和/或加工过的二聚体；或(e) a dimerized, associated and/or processed dimer of CD38 with a soluble ligand such as CD31; or

(f)CD38的一部分。对于多于一种的CD38和/或CD38一部分，例如，CD38特定区域的抗体结合性质在其片段中仍有保留，竞争也存在于CD38BPs间，在这种情况下，位于多个检测片段上的完全呈递的线性抗原决定簇或构象抗原决定簇，与在CD38中一样，呈递在足够大的CD38片段上。(f) A portion of CD38. For more than one type of CD38 and/or part of CD38, for example, the antibody binding properties of a specific region of CD38 are retained in its fragments, competition also exists between CD38BPs, in this case, located on multiple detection fragments Fully presented linear epitopes or conformational epitopes, as in CD38, are presented on sufficiently large fragments of CD38.

典型地，检测竞争性涉及使用第一个量的第一分子；第二个量的第二分子和第三个量的第三分子(或通过结合研究测定的标准，它与以第一和第二分子作为实际同期数据的替代数据的新结合数据相比较更为合理)来评估相对抑制的结合，其中第一、第二和第三个量都是足够的，可以比较，从而给出所讨论的分子与其它存在的分子关于选择性和/特异性方面的信息。第一、第二和第三个量可根据CD38BP的性质和所讨论的潜在靶标有所不同。例如，对于ELISA检验而言，与实施例部分所述类似，需要约5-50μg(例如，约10-50μg、约20-50μg、约5-20μg、约10-20μg等)CD38BP和/或CD38靶标检验是否存在竞争。条件也应该适于结合。典型地，生理或接近生理条件(例如，温度约20-40℃， pH约7-8等)适于CD38BP:CD38结合。Typically, detection of competition involves the use of a first amount of a first molecule; a second amount of a second molecule and a third amount of a third molecule (or a standard determined by binding studies that compares with the first and second It is more reasonable to compare the new binding data of two molecules as a surrogate data for the actual contemporaneous data) to assess the relative inhibition of binding, where the first, second and third quantities are sufficient and can be compared to give the discussed Information about the selectivity and/or specificity of the molecule and other molecules present. The first, second and third amounts may vary depending on the nature of the CD38BP and the potential target in question. For example, for an ELISA assay, about 5-50 μg (e.g., about 10-50 μg, about 20-50 μg, about 5-20 μg, about 10-20 μg, etc.) of CD38BP and/or CD38 is required, similar to that described in the Examples section Target test for competition. Conditions should also be suitable for binding. Typically, physiological or near-physiological conditions (eg, temperature about 20-40° C., pH about 7-8, etc.) are suitable for CD38BP:CD38 binding.

通常，竞争是以通过ELISA和/或FACS分析测定，相对抑制显著高于约5％为特征。需要设定较高的相对抑制的门槛作为特定语境中关于竞争的适当水平的标准/测定值(例如，当竞争分析是用于选择或筛选通过阻断另一种与CD38结合的肽或分子(例如诸如CD31这样的，也称为CD31抗原的CD38天然结合伴侣、EndoCAM、GPIIA1、PECAM-1、血小板/内皮细胞黏附分子或天然存在的抗CD38抗体)的结合功能所设计的新抗体时)。因此，例如，可以设定竞争的标准，其中在认为抗体具有充分竞争性之前，可检测至少约10％的相对抑制，可检测至少约15％的相对抑制，或者可检测至少约20％的相对抑制。在其中属于竞争性抗体的抗原决定簇紧邻抗原的情况下，竞争可为高于40％的CD38结合的相对抑制(例如，至少约45％的抑制，譬如约50％的抑制、例如至少约55％的抑制、譬如约60％的抑制、例如至少约65％的抑制、譬如约70％的抑制、例如至少约75％的抑制、譬如约80％的抑制、例如至少约85％的抑制、譬如约90％的抑制、例如至少约95％的抑制或更高水平的相对抑制)。Typically, competition is characterized by relative inhibition significantly greater than about 5%, as determined by ELISA and/or FACS analysis. A higher threshold of relative inhibition needs to be set as a criterion/determination of an appropriate level of competition in a particular context (e.g. when competition assays are used to select or screen for binding to CD38 by blocking another peptide or molecule (e.g. CD38 natural binding partners such as CD31, also known as CD31 antigen, EndoCAM, GPIIA1, PECAM-1, platelet/endothelial cell adhesion molecule or naturally occurring anti-CD38 antibodies when designing new antibodies) . Thus, for example, criteria for competition can be set wherein a relative inhibition of at least about 10% is detectable, a relative inhibition of at least about 15% is detectable, or a relative inhibition of at least about 20% is detectable before the antibody is considered sufficiently competitive. inhibition. In cases where the antigenic determinant belonging to the competing antibody is in the immediate vicinity of the antigen, the competition may be a relative inhibition of CD38 binding of greater than 40% (e.g., at least about 45% inhibition, such as about 50% inhibition, such as at least about 55% inhibition). % inhibition, such as about 60% inhibition, such as at least about 65% inhibition, such as about 70% inhibition, such as at least about 75% inhibition, such as about 80% inhibition, such as at least about 85% inhibition, such as about 90% inhibition, such as at least about 95% inhibition or a higher level of relative inhibition).

竞争可被认为是某分子和两种潜在结合伴侣之间交叉反应性的另一面。在某些技术方案中，本发明的CD38BP特异地结合到CD38的一个或多个残基或者区域上，但也不与其它肽、肽区域或分子发生交叉反应，例如，本发明提供一种不与诸如BST-1(骨髓基质细胞抗原-1)和也称为CD157的Mo5这样的与CD38同源的蛋白发生交叉反应的抗CD38抗体；或者不与诸如参与多发性骨髓瘤的组织这样的正常组织中CD38发生交叉反应的抗CD38抗体。典型地，没有交叉反应性是指当使用足够量的分子在合适的检测条件下通过ELISA和/或FACS分析进行评估时，在分子间的相对竞争抑制性低于5％。Competition can be considered the flip side of cross-reactivity between a molecule and two potential binding partners. In certain technical schemes, the CD38BP of the present invention specifically binds to one or more residues or regions of CD38, but does not cross-react with other peptides, peptide regions or molecules. For example, the present invention provides a Anti-CD38 antibodies that cross-react with proteins homologous to CD38 such as BST-1 (bone marrow stromal cell antigen-1) and Mo5, also known as CD157; or that do not interact with normal tissues such as those involved in multiple myeloma Anti-CD38 antibodies that cross-react with CD38 in tissues. Typically, no cross-reactivity means a relative competitive inhibition between molecules of less than 5% when assessed by ELISA and/or FACS analysis using sufficient quantities of the molecule under appropriate detection conditions.

在一个技术方案中，本发明提供可与诸如抗体-003这样的具有如SEQ ID No：2中的V_L序列和如SEQ ID No：7中的V_H序列的抗体竞争结合CD38或其部分的CD38BP。In one technical solution, the present invention provides an antibody that can compete with an antibody having a V _L sequence as in SEQ ID No: 2 and a V _H sequence as in SEQ ID No: 7 for binding to CD38 or a portion thereof, such as Antibody-003. CD38BP.

在一个技术方案中，本发明提供可与诸如抗体-005这样的具有如SEQ ID No：12中的V_L序列和如SEQ ID No：17中的V_H序列的抗体竞争结合CD38或其部分的CD38BP。In one technical solution, the present invention provides an antibody that can compete with an antibody having a _VL sequence as in SEQ ID No: 12 and a _VH sequence as in SEQ ID No: 17 for binding to CD38 or a portion thereof, such as Antibody-005. CD38BP.

在一个技术方案中，本发明提供可与诸如抗体-024这样的具有如 SEQ ID No：22中的V_L序列和如SEQ ID No：27中的V_H序列的抗体竞争结合CD38或其部分的CD38BP。In one technical solution, the present invention provides an antibody that can compete for binding to CD38 or a portion thereof with an antibody having a _VL sequence as in SEQ ID No: 22 and a _VH sequence as in SEQ ID No: 27, such as Antibody-024. CD38BP.

如本文其它地方所述，除非特别说明或与内容明显矛盾，涉及CD38BP与CD38的结合是指在任何合适情况下的结合，例如在存在CD38结构的构象环境中；或在线性抗原决定簇环境中。显然，在这种环境的有限子集中的结合是本发明的提供的任何CD38BP的一个重要特性。As described elsewhere herein, reference to CD38BP binding to CD38 refers to binding in any suitable context, such as in a conformational environment where the CD38 structure exists; or in the context of a linear epitope, unless otherwise specified or clearly contradicted by the context. . Clearly, binding in a limited subset of this environment is an important property of any CD38BP provided by the invention.

可在例如Harlow et al.，Antibodies：A Laboratory Manual，ColdSpringHarbor Laboratory Press，Cold Spring Harbor，N.Y.，1988)，Colligan etal.，eds.，Current Protocols in Immunology，Greene Publishing Assoc，andWileyInterScience N.Y.，(1992，1993)，and Muller，Meth.Enzymol.92，589-601(1983))中找到其它通过竞争抑制来测定CD3 8BP特异性的方法。Available, for example, in Harlow et al., Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1988), Colligan et al., eds., Current Protocols in Immunology, Greene Publishing Assoc, and Wiley InterScience N.Y., (1992, 1993 ), and Muller, Meth. Enzymol. 92, 589-601 (1983)) find other methods for determining CD3 8BP specificity by competitive inhibition.

人CD38含有多个不同的抗原决定簇，它们包括(1)包括在人CD38单个肽链中的肽抗原决定簇；(2)由特定链上一个或多个非连续氨基酸和/或在空间上连续，但在不同肽链上(典型地，其中链的各自氨基酸序列在人CD38多肽序列中是不连续的)的氨基酸组成的构象抗原决定簇；(3)由诸如碳水化合物基团这样的全部或部分共价连接到人CD38的分子结构所组成的翻译后抗原决定簇；或(4)(1)-(3)的组合。Human CD38 contains a plurality of different epitopes, which include (1) peptide epitopes included in a single peptide chain of human CD38; A conformational epitope consisting of amino acids that are contiguous, but on different peptide chains (typically, the respective amino acid sequences of the chains are discontinuous in the human CD38 polypeptide sequence); (3) consist of all such as carbohydrate groups Or a post-translational epitope composed of a molecular structure partially covalently linked to human CD38; or a combination of (4)(1)-(3).

本发明中的抗原决定簇包括任何可特异结合到免疫球蛋白上的肽或肽衍生的决定簇。抗原决定簇可在任何位置(对CD38线性序列而言)、方向(对折叠的CD38或其片段而言)包含任何合适数目的氨基酸、氨基酸组合物(因此，至少部分带电)。An antigenic determinant in the present invention includes any peptide or peptide-derived determinant that specifically binds to an immunoglobulin. An antigenic determinant may comprise any suitable number, amino acid composition (thus, at least partially charged) of amino acids in any position (for CD38 linear sequence), orientation (for folded CD38 or fragments thereof).

因此，例如，抗原决定簇可在CD38一级序列的一个或多个连续或不连续位置上含有约3-10个氨基酸，典型地为3-8个氨基酸(例如，抗原决定簇可实质上由分布在CD38中1、2、3、4或5个非连续位置上的2、3、4、5、6、7或8个氨基酸残基组成)。选择性地，例如，可认为抗原决定簇被CD38上约5-40个连续的氨基酸残基的区域所限定(例如，约7-30氨基酸残基、约5-20氨基酸残基或约3-15氨基酸残基)(单独或与相邻的CD38结构域的一部分组合)。在某些抗原决定簇中，可能是只有一个氨基酸残基或仅有几个氨基酸残基对CDR或CDR(s)识别是重要的(因此，对CD38BP:CD38抗原亲和性和强度是最重要的)。 所以，可基于一个或多个这种关键残基来描述抗原决定簇，同时意识到其它残基也对抗原决定簇具有较小的贡献。在通过氨基酸区域来定义抗原决定簇的情况中，在区域中的一个或多个氨基酸对于抗体结合仅有很小的贡献，甚至没有贡献，因此这种残基可用合适的不同残基来替代，并不会导致抗原决定簇对至少某些特异针对它的CD38BPs出现“丢失”。Thus, for example, an antigenic determinant may comprise about 3-10 amino acids, typically 3-8 amino acids, at one or more consecutive or discontinuous positions in the primary sequence of CD38 (e.g., an antigenic determinant may consist essentially of 2, 3, 4, 5, 6, 7 or 8 amino acid residues distributed at 1, 2, 3, 4 or 5 non-contiguous positions in CD38). Alternatively, for example, an antigenic determinant can be considered to be defined by a region of about 5-40 contiguous amino acid residues on CD38 (e.g., about 7-30 amino acid residues, about 5-20 amino acid residues, or about 3- 15 amino acid residues) (alone or in combination with a portion of the adjacent CD38 domain). In some epitopes, it may be that only one amino acid residue or only a few amino acid residues are important for CDR or CDR(s) recognition (thus, for CD38BP: CD38 antigen affinity and strength are the most important of). Therefore, an epitope can be described based on one or more of these key residues, while recognizing that other residues also make minor contributions to the epitope. In cases where the epitope is defined by a region of amino acids, one or more amino acids in the region contribute little or even no contribution to antibody binding, so such residues may be replaced by suitable different residues, does not result in a "loss" of the epitope for at least some of the CD38BPs specific for it.

在一个技术方案中，本发明提供诸如抗CD38抗体这样的CD38BP，它可特异结合到CD38抗原决定簇上，该抗原决定簇也可以被具有如SEQ ID No：2中的V_L序列和如SEQ IDNo：7中的V_H序列的抗体(譬如抗体-003)、或具有如SEQ ID No：12中的V_L序列和如SEQ IDNo：17中的V_H序列的抗体(譬如抗体-005)、或具有如SEQ ID No：22中的V_L序列和如SEQ IDNo：27中的V_H序列的抗体(譬如抗体-024)特异结合。具有一个或多个不同于具有如SEQ IDNo：2中的V_L序列和如SEQ ID No：7中的V_H序列的抗体的CDRs、或具有如SEQ ID No：12中的V_L序列和如SEQ ID No：17中的V_H序列的抗体的CDRs、或具有如SEQ ID No：22中的V_L序列和如SEQ ID No：27中的V_H序列的抗体的CDRs的CDRs的CD38BPs可特异针对分别与作为具有如SEQ IDNo：2中的V_L序列和如SEQ ID No：7中的V_H序列的抗体、或具有如SEQ ID No：12中的V_L序列和如SEQ ID No：17中的V_H序列的抗体、或具有如SEQ ID No：22中的V_L序列和如SEQ IDNo：27中的V_H序列的抗体相同的抗原决定簇。在某些情况下，所述CD3 8BP可识别或比分别具有如SEQ ID No：2中的V_L序列和如SEQ ID No：7中的V_H序列的抗体、或具有如SEQ ID No：12中的V_L序列和如SEQ ID No：17中的V_H序列的抗体、或具有如SEQ ID No：22中的V_L序列和如SEQ ID No：27中的V_H序列的抗体更特异/选择性地针对抗原决定簇的特定结构或区域。In one technical scheme, the present invention provides CD38BP such as anti-CD38 antibody, it can specifically bind on the CD38 epitope, and this epitope can also have as SEQ ID No: V _L sequence in 2 and such as SEQ ID NO: An antibody with a _VH sequence in IDNo:7 (such as antibody-003), or an antibody with a _VL sequence as in SEQ ID No:12 and a _VH sequence as in SEQ IDNo:17 (such as antibody-005), Or an antibody (such as Antibody-024) having a _VL sequence as in SEQ ID No: 22 and a _VH sequence as in SEQ ID No: 27 specifically binds. Having one or more CDRs different from an antibody having a V _L sequence such as SEQ ID No: 2 and a V _H sequence such as SEQ ID No: 7, or having a V _L sequence such as SEQ ID No: 12 and such as The CDRs of the antibody with the _VH sequence in SEQ ID No: 17, or the CD38BPs having the CDRs of the CDRs of the antibody with the _VL sequence in SEQ ID No: 22 and the _VH sequence in SEQ ID No: 27 can be specific Respectively against the antibody having the V _L sequence in SEQ ID No: 2 and the V _H sequence in SEQ ID No: 7, or having the V _L sequence in SEQ ID No: 12 and the antibody in SEQ ID No: 17 The antibody with the _VH sequence in SEQ ID No: 22, or the antibody with the _VL sequence in SEQ ID No: 22 and the _VH sequence in SEQ ID No: 27 has the same epitope. In some cases, the CD3 8BP can recognize or compare with antibodies having a V _L sequence as in SEQ ID No: 2 and a V _H sequence as in SEQ ID No: 7, or an antibody having a sequence as in SEQ ID No: 12 The antibody with the V _L sequence in and the V _H sequence in SEQ ID No: 17, or the antibody with the V _L sequence in SEQ ID No: 22 and the V _H sequence in SEQ ID No: 27 is more specific/ Selectively target specific structures or regions of antigenic determinants.

结合了具有如SEQ ID No：2中的V_L序列和如SEQ ID No：7中的V_H序列的抗体(譬如抗体-003)、或具有如SEQ ID No：12中的V_L序列和如SEQ ID No：17中的V_H序列的抗体(譬如抗体-005)、或具有如SEQ ID No：22中的V_L序列和如SEQ ID No：27中的V_H序列的抗体(譬如抗体-024)的CD38抗原决定簇可通过标准图谱和描述技术来鉴定，可通过合适的技术来鉴定进一步的细节，技术人员可使用这种技术 的多种例子。通常这些技术也可用于鉴定和/或描述针对CD38BPs的抗原决定簇。对这种图谱/描述方法的实施例而言，针对抗CD38抗体的抗原决定簇可通过抗原决定簇的“足迹”对CD38蛋白中暴露的氨基/羧基进行化学修饰的方法来测定。这种足迹技术的一个具体实施例是使用HXMS(通过质谱检测的氢-氘交换)，其中发生了受体和配体蛋白氨基质子的氢/氘交换、结合和回复交换，其中参与蛋白结合的骨架氨基基团在回复交换中受到保护，因此仍为含氘的。在该处通过肽裂解、快速混合高效液相色谱分离和/或电喷雾电离质谱来鉴定相关区域。参见例如Ehring H，AnalyticalBiochemistry，267(2)252-259(1999)and/or Engen，J.R.and Smith，D.L.(2001)Anal.Chem.73，256A-265A。另一个合适的抗原决定簇鉴定技术的实施例是核磁共振抗原决定簇图谱(NMR)，其中典型地，对游离抗原和与诸如抗体这样的抗原结合肽复合在一起的抗原的二维NMR图谱中的信号位置进行比较。典型地，抗原选择性地用同位素¹⁵N标记，这样在NMR图谱上仅有与抗原相关的信号是可见的，而结合了肽的抗原没有信号。典型地，与游离抗原的图谱相比较，来源于氨基酸并参与抗原结合肽相互作用的抗原信号在复合物的图谱有位移，参与结合的氨基酸可被鉴定出来。参见例如Ernst Schering ResFound Workshop.(44)，149-67(2004)、Huang et al.，Journal of MolecularBiology 281(1)，61-67(1998)and Saito and Patterson，Methods.9(3)，516-24(1996)。An antibody (such as Antibody-003) having a _VL sequence as in SEQ ID No: 2 and a _VH sequence as in SEQ ID No: 7, or having a _VL sequence as in SEQ ID No: 12 and a VH sequence as in SEQ ID No: 12 and such as An antibody with a _VH sequence in SEQ ID No: 17 (such as antibody-005), or an antibody with a _VL sequence in SEQ ID No: 22 and a _VH sequence in SEQ ID No: 27 (such as antibody- 024) CD38 epitopes can be identified by standard mapping and delineation techniques, further details can be identified by suitable techniques, of which various examples are available to the skilled artisan. Often these techniques can also be used to identify and/or characterize epitopes directed against CD38BPs. For an example of such a mapping/characterization method, epitopes directed against anti-CD38 antibodies can be determined by chemical modification of exposed amino/carboxyl groups in the CD38 protein by the "footprint" of the epitope. A specific example of this footprinting technique is the use of HXMS (hydrogen-deuterium exchange detected by mass spectrometry), where hydrogen/deuterium exchange, binding and back-exchange of receptor and ligand protein amino protons occur, where the proteins involved in protein binding The backbone amino group is protected in the back exchange and thus remains deuterium-containing. At this point, regions of interest are identified by peptide cleavage, fast-mix HPLC separation, and/or electrospray ionization mass spectrometry. See eg Ehring H, Analytical Biochemistry, 267(2) 252-259 (1999) and/or Engen, JRand Smith, DL (2001) Anal. Chem. 73, 256A-265A. Another example of a suitable epitope identification technique is nuclear magnetic resonance epitope mapping (NMR), where typically two-dimensional NMR spectra of free antigen and antigen complexed with an antigen-binding peptide such as an antibody The position of the signal is compared. Typically, the antigen is selectively labeled with the isotope ¹⁵ N, so that only the signal associated with the antigen is visible on the NMR spectrum, while the peptide-bound antigen has no signal. Typically, the antigen signal derived from the amino acids involved in the interaction of the antigen-binding peptide is shifted in the profile of the complex compared to the profile of the free antigen, and the amino acids involved in the binding can be identified. See, eg, Ernst Schering ResFound Workshop. (44), 149-67 (2004), Huang et al., Journal of Molecular Biology 281(1), 61-67 (1998) and Saito and Patterson, Methods. 9(3), 516 -24 (1996).

也可用质谱方法进行抗原决定簇绘图/描述。参见，例如Downward，J MassSpectrom.35(4)，493-503(2000)and Kiselar and Downard，Anal Chem.71.(9)，1792-801(1999)。Epitope mapping/description can also be performed using mass spectrometry methods. See, eg Downward, J Mass Spectrom. 35(4), 493-503 (2000) and Kiselar and Downard, Anal Chem. 71.(9), 1792-801 (1999).

蛋白酶消化技术也可用于抗原决定簇绘图和鉴定。可通过蛋白酶消化测定抗原决定簇相关的区域/序列，例如，通过胰蛋白酶以与CD38约1∶50的比例，在37℃，pH7-8，过夜(O/N)消化，然后通过质谱(MS)分析进行肽鉴定。然后可通过比较进行胰蛋白酶消化的样品和CD38BP温育后，再进行例如胰蛋白酶消化的样品，可鉴定出在胰蛋白酶切割中被CD38BP保护的肽(从而表明结合伴侣的足迹)。其它的胰凝乳蛋白酶、胃蛋白酶等酶也可或选择性地用作同样的抗原决定簇描述方法。在这些检测中，与具有如SEQ ID No：2中的V_L序列和如SEQ ID No：7中的V_H序列的抗体(譬如抗体-003)、或具有如SEQ ID No：12中的 V_L序列和如SEQ ID No：17中的V_H序列的抗体(譬如抗体-005)、或具有如SEQ ID No：22中的V_L序列和如SEQ ID No：27中的V_H序列的抗体(譬如抗体-024)具有显著相同的结果的CD3 8BP可被认为是与分别具有如SEQ ID No：2中的V_L序列和如SEQ ID No：7中的V_H序列的抗体(譬如抗体-003)、或具有如SEQ ID No：12中的V_L序列和如SEQID No：17中的V_H序列的抗体(譬如抗体-005)、或具有如SEQ ID No：22中的V_L序列和如SEQ ID No：27中的V_H序列的抗体(譬如抗体-024)结合相同抗原决定簇的抗体。例如，参见Manca，Ann 1st Super Sanita.27(1)，15-9(1991)讨论的类似技术。通过其中一个抗体是生物素化的两个抗体与CD38竞争结合而获得的抗原决定簇图谱是鉴定相关抗原决定簇区域的另一种方法。Protease digestion techniques can also be used for epitope mapping and identification. The region/sequence associated with the epitope can be determined by protease digestion, e.g., by trypsin at a ratio of approximately 1:50 to CD38, overnight (O/N) at 37°C, pH 7-8, followed by mass spectrometry (MS ) analysis for peptide identification. Peptides that are protected by CD38BP during trypsin cleavage (thus indicating the footprint of the binding partner) can then be identified by comparing trypsinized samples with CD38BP-incubated, eg trypsinized samples. Other enzymes such as chymotrypsin, pepsin, etc. can also or alternatively be used for the same epitope delineation method. In these assays, an antibody (such as antibody-003) having a V _L sequence as in SEQ ID No: 2 and a V _H sequence as in SEQ ID No: 7, or having a V as in SEQ ID No: 12 An antibody with a _L sequence and a _VH sequence as in SEQ ID No: 17 (such as Antibody-005), or an antibody with a _VL sequence as in SEQ ID No: 22 and a _VH sequence as in SEQ ID No: 27 (such as Antibody-024) _{CD3 8BP} having significantly the same result can be considered as the antibody (such as antibody- 003), or have the _VL sequence as in SEQ ID No: 12 and the antibody (such as antibody-005) as the _VH sequence in SEQID No: 17, or have the _VL sequence as in SEQ ID No: 22 and An antibody with a _VH sequence as in SEQ ID No: 27 (eg Antibody-024) binds an antibody to the same epitope. See, eg, a similar technique discussed in Manca, Ann 1st Super Sanita. 27(1), 15-9 (1991). Epitope mapping by competitive binding of two antibodies to CD38, one of which is biotinylated, is another method for identifying regions of related epitopes.

通过基于PEPSCAN的酶联免疫方法来检测抗体与线状或环状CD38肽的结合是另一种鉴定相关抗原决定簇区域的方法，例如，参见Slootstra-JW et al.Mol-Divers.1，87-96(1996)。定点突变是鉴定相关抗原决定簇区域的另一种方法，参见，例如Polyak andDeans，Blood 99，3956-3962(2002)。Detection of antibody binding to linear or cyclic CD38 peptides by PEPSCAN-based ELISA is another way to identify relevant epitopic regions, see for example Slootstra-JW et al. Mol-Divers.1, 87 -96 (1996). Site-directed mutagenesis is another approach to identify regions of relevant epitopes, see eg Polyak and Deans, Blood 99, 3956-3962 (2002).

各种噬菌体展示技术也可用于鉴定抗原决定簇。参见，例如Wangand Yu，CurrDrug Targets.5(1)，1-15(2004)、Burton，Immunotechnology.1(2)，87-94(1995 Aug)，Cortese et al.，Immunotechnology.1(2)，87-94(1995)和Irving et al.，Curr OpinChem Biol.5(3)，314-24(2001)。一致的抗原决定簇也可通过修改的噬菌体展示相关的技术进行鉴定(参见http://www.cs.montana.edu/～mumey/papers/jcb03.pdf)。Various phage display techniques can also be used to identify epitopes. See, for example, Wang and Yu, CurrDrug Targets. 5(1), 1-15 (2004), Burton, Immunotechnology. 1(2), 87-94 (1995 Aug), Cortese et al., Immunotechnology. 1(2), 87-94 (1995) and Irving et al., Curr Opin Chem Biol. 5(3), 314-24 (2001). Consensus epitopes can also be identified by a modified phage display-related technique (see http://www.cs.montana.edu/~mumey/papers/jcb03.pdf ).

其它可能进行抗原决定簇绘图的方法包括晶体技术、X-射线衍射技术(譬如由Poljak和其它人在1970s-1980s开发的X-射线衍射/序列研究技)、和多中心肽合成的应用。诸如序列分析和三维结构分析等基于计算机的方法也可用于鉴定抗原决定簇。例如，可通过CD38的结构和进入的单个单克隆抗体Fab片段的结构的分子建模来测定抗原决定簇。这些和其它绘图方法在Epitope Mapping A Practical Approach(Westwoodand Hay Eds.)2001 Oxford University Press中进行了讨论。Other possible methods for epitope mapping include crystallographic techniques, X-ray diffraction techniques (such as X-ray diffraction/sequencing techniques developed by Poljak and others in the 1970s-1980s), and the use of polycentric peptide synthesis. Computer-based methods such as sequence analysis and three-dimensional structure analysis can also be used to identify epitopes. For example, epitopes can be determined by molecular modeling of the structure of CD38 and the structure of incoming single monoclonal antibody Fab fragments. These and other mapping methods are discussed in Epitope Mapping A Practical Approach (Westwood and Hay Eds.) 2001 Oxford University Press.

在一个技术方案中，本发明提供实质具有一个或多个选自具有如SEQ ID No：2中的V_L序列和如SEQ ID No：7中的V_H序列的抗体(譬如抗体-003)、或具有如SEQ ID No：12中的V_L序列和如SEQ ID No：17中的V_H序列的抗体(譬如抗体-005)、或具有如SEQ ID No：22中的V_L序列和如SEQ ID No：27中的V_H序列的抗体(譬如抗体-024)的mAbs的相同的特异CD38结合特征的CD38BP。In one technical solution, the present invention provides essentially having one or more antibodies (such as antibody-003) selected from the group consisting of _VL sequences such as SEQ ID No: 2 and _VH sequences such as SEQ ID No: 7, Or have the _VL sequence as in SEQ ID No: 12 and the antibody (for example antibody-005) as the _VH sequence in SEQ ID No: 17, or have the _VL sequence as in SEQ ID No: 22 and as SEQ ID No: CD38BP with the same specific CD38 binding characteristics as mAbs of antibodies with _VH sequences in ID No: 27 (such as Antibody-024).

绘图研究已表明，几种增强针对人CD38的单克隆抗体是与CD38的C末端区域的抗原决定簇(220-296)结合的(Hoshino et al.and Ferrero etal.)。在人和猕猴CD38序列间在该区域有3个氨基酸差异：在人中的T237、Q272和S274对应于猕猴中的A238、R273和F275。-005不与猕猴组织结合(如实施例10和11所示)。人和猴CD38序列存在有限数目的氨基酸差异，例如蛋白的羧基末端部分，例如以下人和猕猴CD38序列间的3个氨基酸差异，在人CD38s中的T237、Q272和S274对应于猕猴的猴CD38中的A238、R273和F275(比较SEQ IDNo.21和SEQ ID No.22)。-005与突变的其中SEQ ID No：31的272位谷氨酰胺残基被精氨酸残基替代(Q272R)人CD38蛋白、或突变的其中SEQID No：31的274位丝氨酸残基被苯丙氨酸残基替代(S274F)(如实施例17所示)人CD38蛋白的结合程度与其和野生型人CD38的结合程度不同。特别地，与-005的结合可被S274F位置的氨基酸替代所消除。Mapping studies have shown that several monoclonal antibodies enhanced against human CD38 bind to epitopes (220-296) in the C-terminal region of CD38 (Hoshino et al. and Ferrero et al.). There are 3 amino acid differences in this region between human and macaque CD38 sequences: T237, Q272 and S274 in human correspond to A238, R273 and F275 in macaque. -005 did not bind to macaque tissue (as shown in Examples 10 and 11). There are a limited number of amino acid differences between human and monkey CD38 sequences, such as the carboxy-terminal part of the protein, for example the following 3 amino acid differences between human and macaque CD38 sequences, T237, Q272 and S274 in human CD38s correspond to monkey CD38 in macaques A238, R273 and F275 (compare SEQ ID No. 21 and SEQ ID No. 22). -005 and mutated wherein SEQ ID No: 272 glutamine residues of 31 are replaced by arginine residues (Q272R) human CD38 protein, or mutated wherein SEQ ID No: 274 serine residues of 31 are replaced by phenylpropanoids Amino acid residue substitution (S274F) (as shown in Example 17) has a different binding degree to human CD38 protein than to wild-type human CD38. In particular, binding to -005 was abrogated by an amino acid substitution at position S274F.

因此，本发明提供与人CD38(SEQ ID No：31)结合的肽，它与其中272位谷氨酰胺残基被精氨酸残基替代的、突变的人CD38(SEQ IDNo：33)的结合程度不同于与其与人CD38(SEQ ID No：31)的结合程度。Accordingly, the present invention provides peptides that bind to human CD38 (SEQ ID No: 31), which bind to mutated human CD38 (SEQ ID No: 33) in which the glutamine residue at position 272 is replaced by an arginine residue. The extent differs from its binding to human CD38 (SEQ ID No: 31).

本发明也提供与人CD38(SEQ ID No：31)结合的肽，它与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ IDNo：34)的结合程度不同于与其与人CD38(SEQID No：31)的结合程度。The present invention also provides peptides that bind to human CD38 (SEQ ID No: 31) to a different extent than mutated human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is replaced by a phenylalanine residue in the degree of its binding to human CD38 (SEQ ID No: 31).

术语“相同程度”应理解为，肽与突变人CD38的结合显著低于肽与野生型人CD38的结合。肽与CD38分子(野生型和突变型)的结合可通过各种本领域技术人员熟知的方法来测定与突变型的结合是否“显著低于”与野生型的结合。本领域技术人员可使用各种测定肽与另一种肽的结合的不同技术，例如ELISA、放射性免疫检测、BIAcore或流式细胞仪。The term "to the same extent" is understood to mean that the binding of the peptide to mutant human CD38 is significantly lower than the binding of the peptide to wild-type human CD38. Binding of peptides to CD38 molecules (wild-type and mutant) can be determined by various methods well known to those skilled in the art to determine whether binding to the mutant is "significantly lower" than binding to the wild-type. Various techniques for determining the binding of a peptide to another peptide are available to those skilled in the art, such as ELISA, radioimmunoassay, BIAcore or flow cytometry.

一个测定结合的方法是通过测定肽与突变蛋白的结合的EC₅₀，然后比较所获得的数值。另一种测定结合的方法是通过检测在饱和浓度结合的数量(例如，结合信号的平台)，或通过诸如BIAcore来测定动力学速率常数k_on和k_off。One method of determining binding is by determining the _EC50 of the binding of the peptide to the mutein and then comparing the values obtained. Another way to measure binding is by measuring the amount bound at saturating concentrations (eg, the plateau of the binding signal), or by determining kinetic rate constants _kon and _koff such as by BIAcore.

在一个技术方案中，所述肽与CD38蛋白(突变型或野生型)的结合通过如实施例17所述的ELISA来测定。In one embodiment, the binding of the peptide to CD38 protein (mutant or wild type) is determined by ELISA as described in Example 17.

在一个技术方案中，肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的50％。在一个技术方案中，肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的10％。在一个技术方案中，肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的5％。在一个技术方案中，肽与其中274位丝氨酸残基被苯丙氨酸残基替代的、突变的人CD38(SEQ ID No：34)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的1％。In one embodiment, the _EC50 of the peptide binding to mutant human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is replaced by a phenylalanine residue is less than that of the peptide binding to human CD38 (SEQ ID No: 34). :31) ₅₀ % of the EC50 of binding. In one embodiment, the _EC50 of the peptide binding to mutated human CD38 (SEQ ID No: 34) in which the 274th serine residue is replaced by a phenylalanine residue is less than that of the peptide binding to human CD38 (SEQ ID No: 31) 10% of the _EC50 of binding. In one embodiment, the _EC50 of the peptide binding to mutant human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is replaced by a phenylalanine residue is less than that of the peptide binding to human CD38 (SEQ ID No: 34). :31) 5% of the _EC50 of binding. In one embodiment, the _EC50 of the peptide binding to mutant human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is replaced by a phenylalanine residue is less than that of the peptide binding to human CD38 (SEQ ID No: 34). :31) 1% of the _EC50 of binding.

在一个技术方案中，肽与其中272位谷氨酰胺残基被精氨酸残基替代的、突变的人CD38(SEQ ID No：33)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的50％。在一个技术方案中，肽与其中272位谷氨酰胺残基被精氨酸残基替代的、突变的人CD38(SEQID No：33)结合的EC₅₀不到该肽与人CD38(SEQ ID No：31)结合的EC₅₀的10％。In one embodiment, the _EC50 of the peptide binding to mutant human CD38 (SEQ ID No: 33) in which the glutamine residue at position 272 is replaced by an arginine residue is less than that of the peptide binding to human CD38 (SEQ ID NO: 33) No: 31) ₅₀ % of the EC50 of binding. In one embodiment, the _EC50 of the peptide binding to mutant human CD38 (SEQ ID No: 33) in which the glutamine residue at position 272 is replaced by an arginine residue is less than that of the peptide binding to human CD38 (SEQ ID No: 33). :31) 10% of the _EC50 of binding.

在一个技术方案中，肽与其中237位苏氨酸残基被丙氨酸残基替代的、突变的人CD38(SEQ ID No：32)结合，结合程度与其和人CD38(SEQ ID No：31)结合相同。在一个技术方案中，肽与突变的其中237位苏氨酸残基被丙氨酸残基替代的人CD38结合(SEQ IDNo：32)的EC₅₀大于肽与人CD38(SEQ ID No：31)结合的EC₅₀的75％。在一个技术方案中，肽与突变的其中237位苏氨酸残基被丙氨酸残基替代的人CD38结合(SEQ ID No：32)的EC₅₀大于肽与人CD38(SEQ ID No：31)结合的EC₅₀的85％。在一个技术方案中，肽与突变的其中237位苏氨酸残基被丙氨酸残基替代的人CD38结合(SEQID No：32)的EC₅₀大于肽与人CD38(SEQ ID No：31)结合的EC₅₀的90％。在一个技术方案中，肽与突变的其中237位苏氨酸残基被丙氨酸残基替代的人CD38结合(SEQ ID No：32)的EC₅₀大于肽与人CD38(SEQ ID No：31)结合的EC₅₀的95％。In one embodiment, the peptide binds to the same extent as human CD38 (SEQ ID No: 31 ) combine the same. In one technical solution, the _EC50 of the peptide binding to a mutated human CD38 (SEQ ID No: 32) in which the 237-threonine residue is replaced by an alanine residue is greater than that of the peptide binding to human CD38 (SEQ ID No: 31) 75% of _EC50 for binding. In one technical solution, the _EC50 of the peptide binding to a mutated human CD38 (SEQ ID No: 32) in which the 237th threonine residue is replaced by an alanine residue is greater than that of the peptide binding to human CD38 (SEQ ID No: 31 ) 85% of the _EC50 for binding. In one technical solution, the _EC50 of the peptide binding to a mutated human CD38 (SEQ ID No: 32) in which the 237-threonine residue is replaced by an alanine residue is greater than that of the peptide binding to human CD38 (SEQ ID No: 31) 90% of _EC50 for binding. In one technical solution, the _EC50 of the peptide binding to a mutated human CD38 (SEQ ID No: 32) in which the 237th threonine residue is replaced by an alanine residue is greater than that of the peptide binding to human CD38 (SEQ ID No: 31 ) 95% of the _EC50 for binding.

为了在CD38中鉴定更特异的可能的抗原决定簇区域，可使用各种预测的分析方法。在第一个分析方法中，可分析CD38的(1)高度疏水区域(使用Kyte-Doolittle方法)；(2)通过原外凸区指数方法测定抗原性；(3)通过Parker方法测定抗原性；(4)通过Hopp/Woods方法测定抗原性；及(5)通过Goldman，Engleman，and Steitz方法测定亲水性。可根据具有一个或多个这些特性来选择长10-40个氨基酸的序列。该方法的原则是通用的同一性是：许多理想的B细胞抗原决定簇是长8-10个氨基酸的亲水的、表面定向的及柔性序列。In order to identify more specific regions of potential epitopes in CD38, various predictive assays can be used. In the first assay, CD38 can be analyzed for (1) highly hydrophobic regions (using the Kyte-Doolittle method); (2) antigenicity by the original convex region index method; (3) antigenicity by the Parker method; (4) Antigenicity was determined by the Hopp/Woods method; and (5) Hydrophilicity was determined by the Goldman, Engleman, and Steitz method. Sequences 10-40 amino acids in length can be selected for having one or more of these properties. The principle of this approach is that the general identity is that many ideal B cell epitopes are hydrophilic, surface oriented and flexible sequences 8-10 amino acids long.

本发明提供针对以这种方式鉴定的CD38的CD38区域的CD38BPs。另外，这些序列的末端可与预测的通过此处所述其它分析获得的抗原决定簇的区域，从而提供其它的特定的可能含抗原决定簇的区域。可进行其它类似的比较，从而提供其它的可能的抗原决定簇区域，其中与这些抗原决定簇区域结合的CD38BPs可认为是本发明的另一个特征。The present invention provides CD38BPs directed against the CD38 region of CD38 identified in this manner. In addition, the termini of these sequences can be compared to regions predicted to be epitopes obtained by other assays described herein, thereby providing additional specific regions that may contain epitopes. Other similar comparisons can be made to provide other possible epitopic regions to which CD38BPs bind can be considered as another feature of the invention.

在一个技术方案中，本发明的CD38BP是抗体。本发明提供的CD38结合免疫球蛋白分子的非限定的例子包括(a)含以下组成的有完全功能的免疫球蛋白分子：(i)两个含具有人B细胞表面抗原特异性的可变区和人恒定区的相同的嵌合重链，及(ii)两个相同的全(即，非嵌合)人轻链；(b)含以下组成的有完全功能的免疫球蛋白分子(i)两个含如所述的可变区和人恒定区的嵌合重链，及(ii)两个相同的全(即，非嵌合)非人轻链；(c)单价抗体，即，含以下组成的有完全功能的免疫球蛋白分子(i)两个含如所述的可变区和人恒定区的嵌合重链，就(ii)两个不同的轻链，其中只有一个具有与重链可变区相同的特异性。得到的抗体分子仅与其一个末端结合，而不能进行二价结合。如示例，本发明提供的免疫球蛋白相关的肽可包括以下组成：(a)整个免疫球蛋白分子；(b)scFv；(c)单克隆抗体；(d)人抗体；(e)嵌合抗体；)(f)人源化抗体；(g)Fab片段；(h)Fab1片段；(i)F(ab′) ₂片段；(j)Fv分子；及(k)二硫键连接的Fv分子。In one technical solution, the CD38BP of the present invention is an antibody. Non-limiting examples of CD38-binding immunoglobulin molecules provided by the invention include (a) fully functional immunoglobulin molecules comprising (i) two variable domains specific for human B cell surface antigens A chimeric heavy chain identical to that of a human constant region, and (ii) two identical fully (i.e., non-chimeric) human light chains; (b) a fully functional immunoglobulin molecule comprising (i) Two chimeric heavy chains comprising variable and human constant regions as described, and (ii) two identical full (i.e., non-chimeric) non-human light chains; (c) a monovalent antibody, i.e., comprising A fully functional immunoglobulin molecule consisting of (i) two chimeric heavy chains containing variable and human constant regions as described, and (ii) two different light chains, only one of which has the same Same specificity for the heavy chain variable region. The resulting antibody molecule binds only to one of its ends and cannot undergo bivalent binding. As an example, immunoglobulin-related peptides provided herein may comprise the following compositions: (a) whole immunoglobulin molecules; (b) scFv; (c) monoclonal antibodies; (d) human antibodies; (e) chimeric Antibodies; (f) humanized antibodies; (g) Fab fragments; (h) Fab1 fragments; (i) F(ab') ₂ fragments; (j) Fv molecules; and (k) disulfide-linked Fv molecular.

在一个技术方案中，本发明的CD38BP是多克隆抗体。在一个技术方案中，本发明的CD38BP是单克隆抗体。在另一个技术方案中，本发明的CD38BP是人单克隆抗体。在另一个技术方案中，本发明的CD38BP是人源化抗体。在另一个技术方案中，本发明的CD38BP是嵌合抗体。 在另一个技术方案中，本发明的CD38BP是完全来源于不同于人的哺乳动物物种的单克隆抗体。在另一个技术方案中，本发明的CD38BP是完全的鼠单克隆抗体。In one technical solution, the CD38BP of the present invention is a polyclonal antibody. In one technical solution, the CD38BP of the present invention is a monoclonal antibody. In another technical solution, the CD38BP of the present invention is a human monoclonal antibody. In another technical solution, the CD38BP of the present invention is a humanized antibody. In another technical solution, the CD38BP of the present invention is a chimeric antibody. In another technical solution, the CD38BP of the present invention is a monoclonal antibody derived entirely from a mammalian species other than humans. In another technical solution, the CD38BP of the present invention is a complete mouse monoclonal antibody.

单克隆抗体是指含具有相同结构和特异性的同源抗体群的组分。典型地，单克隆抗体是从实质同源的抗体，即含除了存在微量可能的天然发生的突变外，相同的群组成的单个抗体，获得的抗体。单克隆抗体是高度特异的，每个单克隆抗体典型地针对单个抗原决定簇，相反，多克隆抗体制品典型地包括针对不同抗原决定簇的不同抗体。抗体是单克隆的并不表示需要通过任何特定的方法来产生抗体。例如。本发明的单克隆抗体可通过Kohleret al.，Nature 256，495(1975)描述的杂交方法来产生，或可通过重组DNA方法来产生。单克隆抗体也可使用，例如Clackson et al.，Nature 352，624-628(1991)和Marks et al.，J.MoI.Biol.222.581-597(1991)所述的技术，从噬菌体抗体文库中分离。Monoclonal antibody refers to a composition comprising a population of homologous antibodies having the same structure and specificity. Typically, a monoclonal antibody is one obtained from substantially homologous antibodies, ie, a single antibody comprising the same population except for the presence of minor possible naturally occurring mutations. Monoclonal antibodies are highly specific, with each monoclonal antibody typically directed against a single epitope, in contrast, polyclonal antibody preparations typically include different antibodies directed against different epitopes. The fact that an antibody is monoclonal does not necessarily mean that the antibody was produced by any particular method. E.g. The monoclonal antibodies of the present invention can be produced by the hybridization method described in Kohler et al., Nature 256, 495 (1975), or can be produced by recombinant DNA methods. Monoclonal antibodies can also be obtained from phage antibody libraries using, for example, the techniques described in Clackson et al., Nature 352, 624-628 (1991) and Marks et al., J. MoI. Biol. 222.581-597 (1991). separate.

单克隆抗体可从任何合适的来源获得。因此，例如，单克隆抗体可从用目的抗原，例如，以在表面表达抗原的细胞形式，或者编码目的抗原的核酸形式，所免疫的鼠脾B细胞所制备的杂交瘤中获得。单克隆抗体也可从来源于免疫的人或诸如鼠、狗、灵长类等非人哺乳动物中的抗体表达细胞的杂交瘤中获得。Monoclonal antibodies can be obtained from any suitable source. Thus, for example, monoclonal antibodies can be obtained from hybridomas prepared from murine splenic B cells immunized with the antigen of interest, eg, in the form of cells expressing the antigen on their surface, or in the form of nucleic acids encoding the antigen of interest. Monoclonal antibodies can also be obtained from hybridomas derived from antibody-expressing cells in immunized humans or non-human mammals such as mice, dogs, primates, and the like.

选择性地，可在其它表达系统，包括原核细胞，譬如诸如大肠杆菌这样的微生物、藻类和昆虫细胞中表达克隆的抗体基因，用于产生单链Fv抗体。另外，抗体可在诸如羊和兔奶或鸡蛋这样的转基因非人动物，或在转基因植物中进行生产。残基，例如Verma，R.，etal.，J.lmmunol.Meth.216，165-181(1998)；Pollock，et al.，J.lmmunoi.Meth.231，147-157(1999)；and Fischer，R.，et al.，Biol.Chem.380，825-839(1999)。Alternatively, cloned antibody genes can be expressed in other expression systems, including prokaryotic cells, such as microorganisms such as E. coli, algae and insect cells, for the production of single chain Fv antibodies. In addition, antibodies can be produced in transgenic non-human animals such as goat and rabbit milk or eggs, or in transgenic plants. residues, such as Verma, R., et al., J.lmmunol.Meth.216, 165-181 (1998); Pollock, et al., J.lmmunoi.Meth.231, 147-157 (1999); and Fischer , R., et al., Biol. Chem. 380, 825-839 (1999).

在一个技术方案中，可使用携带部分人免疫系统，而不是鼠系统的转基因或转染色体小鼠来产生直接针对CD38的人单克隆抗体。这种转基因和转染色体小鼠分别包括此处所指的HuMAb鼠和KM鼠，此处统称为“转基因鼠”。在这种鼠中产生的人单克隆抗体简称为HuMab。In one embodiment, transgenic or transchromosomal mice carrying parts of the human immune system, rather than the murine system, can be used to generate human monoclonal antibodies directed against CD38. Such transgenic and transchromosomal mice include HuMAb mice and KM mice referred to herein, respectively, and are collectively referred to as "transgenic mice" herein. The human monoclonal antibody produced in this mouse is called HuMab for short.

HuMAb小鼠含编码未重排人重链(μ和γ)和κ轻链免疫球蛋白序列的人免疫球蛋白具有微座位与内源μ个κ链基因座位失活的目的突变(Lonberg，N.et al.，Nature 368，856-859(1994))。因此，小鼠在 免疫应答时表现出鼠IgM或κ的降低表达，所导入的人重链和轻链转基因经过类别转换和体细胞突变生成高亲和性的人IgG，κ单克隆抗体(Lonberg，N.etal.(1994)、supra；reviewed in Lonberg，N.Handbookof Experimental Pharmacology113，49-101(1994)，Lonberg，N.andHuszar，D.，Intern.ReV.Immunol.Vol.13 65-93(1995)and Harding，F.and Lonberg，N.Ann.N.Y.Acad.Sci 764 536-546(1995))。在Taylor，L.etal.，Nucleic Acids Research 20，6287-6295(1992)、Chen，J.et al.，InternationalImmunology 5，647-656(1993)、Tuaillon et al.，J.Immunol.152，2912-2920(1994)、Taylor，L.et al.，International Immunology 6，579-591(1994)、Fishwild，D.et al.，Nature Biotechnology 14，845-851(1996)中详细描述了HuMAb小鼠的制备。也参见，US 5,545,806、US 5,569,825、US 5,625,126、US 5,633,425、US 5,789,650、US 5,877,397、US5,661,016、US 5,814,318、US 5,874,299、US 5,770,429、US 5,545,807、WO 98/24884、WO94/25585、WO 93/1227、WO 92/22645、WO 92/03918和WO 01/09187。HuMAb mice contain human immunoglobulin sequences encoding unrearranged human heavy (μ and γ) and kappa light chain immunoglobulin sequences with mutations of interest that inactivate the microloci and the endogenous μ kappa chain loci (Lonberg, N et al., Nature 368, 856-859 (1994)). Therefore, mice exhibit reduced expression of murine IgM or κ in response to immune responses, and the introduced human heavy and light chain transgenes undergo class switching and somatic mutation to generate high-affinity human IgG, κ monoclonal antibodies (Lonberg , N. etal. (1994), supra; reviewed in Lonberg, N. Handbook of Experimental Pharmacology 113, 49-101 (1994), Lonberg, N. and Huszar, D., Intern. 1995) and Harding, F. and Lonberg, N. Ann. N. Y. Acad. Sci 764 536-546 (1995)). In Taylor, L. et al., Nucleic Acids Research 20, 6287-6295 (1992), Chen, J. et al., International Immunology 5, 647-656 (1993), Tuaillon et al., J. Immunol.152, 2912 HuMAb mice are described in detail in -2920 (1994), Taylor, L. et al., International Immunology 6, 579-591 (1994), Fishwild, D. et al., Nature Biotechnology 14, 845-851 (1996) preparation.也参见，US 5,545,806、US 5,569,825、US 5,625,126、US 5,633,425、US 5,789,650、US 5,877,397、US5,661,016、US 5,814,318、US 5,874,299、US 5,770,429、US 5,545,807、WO 98/24884、WO94/25585、WO 93/1227 , WO 92/22645, WO 92/03918 and WO 01/09187.

HCo7小鼠在其内源轻链(κ)基因中具有JKD破坏(如Chen et al.，EMBO J.12，821-830(1993)所述)，在其内源重链基因中有CMD破坏(如WO 01/14424中实施例1所述)。KCo5人κ轻链转基因如Fishwildet al.，Nature Biotechnology 14，845-851(1996))所述，HCo7人重链转基因(如US 5,770,429所述)。HCo7 mice have a JKD disruption in their endogenous light chain (κ) gene (as described by Chen et al., EMBO J.12, 821-830 (1993)) and a CMD disruption in their endogenous heavy chain gene (as described in Example 1 of WO 01/14424). KCo5 human kappa light chain transgene as described in Fishwild et al., Nature Biotechnology 14, 845-851 (1996)), HCo7 human heavy chain transgene as described in US 5,770,429.

Hco12小鼠在其内源轻链(κ)基因中具有JKD破坏(如Chen et al.，EMBO J.12，821-830(1993)所述)，在其内源重链基因中有CMD破坏(如WO 01/14424中实施例1所述)。KCo5人κ轻链转基因如Fishwildet al.，Nature Biotechnology 14，845-851(1996))所述，HCo12人重链转基因(如WO 01/14424中实施例2所述)。在KM小鼠株中，已按照Chen et al.，EMBO J.12，811-820(1993)所述对小鼠内源κ轻链基因进行纯合破坏，并按WO 01/09187的实施例1中所述对内源小鼠重链基因进行纯合破坏。该小鼠株携带人κ轻链转基因KCo5，如Fishwild etal.，Nature Biotechnology 14，845-851(1996)所述。该小鼠株也携带由染色体14片段hCF(SC20)组成的人重链转染色体，如WO 02/43478所述。Hco12 mice have a JKD disruption in their endogenous light chain (κ) gene (as described by Chen et al., EMBO J.12, 821-830 (1993)) and a CMD disruption in their endogenous heavy chain gene (as described in Example 1 of WO 01/14424). KCo5 human kappa light chain transgene as described by Fishwild et al., Nature Biotechnology 14, 845-851 (1996)), HCo12 human heavy chain transgene (as described in Example 2 of WO 01/14424). In the KM mouse strain, the mouse endogenous kappa light chain gene has been homozygously disrupted as described by Chen et al., EMBO J.12, 811-820 (1993), and as described in the example of WO 01/09187 Homozygous disruption of the endogenous mouse heavy chain gene was performed as described in 1. This mouse strain carries the human kappa light chain transgene KCo5 as described by Fishwild et al., Nature Biotechnology 14, 845-851 (1996). This mouse strain also carries a human heavy chain transchromosome consisting of the chromosome 14 fragment hCF (SC20), as described in WO 02/43478.

KM小鼠含人重链转染色体和人κ轻链转基因。在KM鼠中，内源 鼠重链和轻链基因也被破坏，这样对鼠免疫可产生人免疫球蛋白，而不是鼠免疫球蛋白。在WO 02/43478中详细描述了KM小鼠的构建及在产生人免疫球蛋白方面的用途。KM mice contain a human heavy chain transchromosome and a human kappa light chain transgene. In KM mice, the endogenous mouse heavy and light chain genes are also disrupted so that immunization of the mice produces human rather than murine immunoglobulins. The construction of KM mice and their use in the production of human immunoglobulins is described in detail in WO 02/43478.

根据众所周知的技术，来自这些转基因小鼠的脾细胞可用于产生能分泌人单克隆抗体的杂交瘤。这些转基因哺乳动物、含编码可操作的核酸序列用于表达CD38BP的哺乳动物、用一个或单个编码CD38核酸序列稳定转染的哺乳动物等是本发明的另外的特征。Splenocytes from these transgenic mice can be used to generate hybridomas secreting human monoclonal antibodies according to well known techniques. These transgenic mammals, mammals containing an operable encoding nucleic acid sequence for expression of CD38BP, mammals stably transfected with one or a single CD38 encoding nucleic acid sequence, etc. are additional features of the invention.

本发明的人单克隆或多克隆抗体，或本发明来源于其它物种的抗体也可通过另一种非人动物或植物后代经转基因方式产生，该非人哺乳动物或植物是用目的免疫球蛋白重链和轻链序列进行转基因的，并能以可回收的形式产生抗体。与哺乳动物中转基因产生相关，抗体可以羊、牛或其它哺乳动物的乳汁的可回收形式来生产。参见，例如，US5,827,690、US 5,756,687、US 5,750,172和US 5,741,957。The human monoclonal or polyclonal antibodies of the present invention, or the antibodies of the present invention derived from other species, can also be produced transgenically by the progeny of another non-human animal or plant using the immunoglobulin of interest The heavy and light chain sequences are transgenic and antibodies can be produced in a retrievable form. In connection with transgene production in mammals, antibodies can be produced in a recoverable form in the milk of sheep, cows or other mammals. See, eg, US 5,827,690, US 5,756,687, US 5,750,172 and US 5,741,957.

此外，本发明的人抗体或本发明来源于其它物种的抗体可使用本领域周知的工艺，通过展示类技术来生产，包括但不限定于，噬菌体展示、逆转录病毒展示、核糖体展示及其它技术，所获得的分子可用于诸如亲和力成熟这样的另外的成熟，这类技术在本领域是周知的(参见，例如Hoogenboom et al.，J.MoI.Biol.227.381(1991)(噬菌体展示)，Vaughanet al.，Nature Biotech 14.309(1996)(噬菌体展示)，Hanes and Plucthau，PNASUSA 94，4937-4942(1997)(核糖体展示)，Parmley and Smith，Gene 73，305-318(1988)(噬菌体展示)，Scott TIBS 17，241-245(1992)、Cwirla et al.，PNAS USA 87，6378-6382(1990)、Russet et al.，Nucl.AcidsResearch 21，1081-1085(1993)、Hoogenboom et al.，Immunol.Reviews130，43-68(1992)、Chiswell and McCafferty TIBTECH 10，80-84(1992)、and US 5,733,743)。如果展示技术用于产生非人抗体，则这类抗体可被人源化，例如本文其它部分所述。In addition, the human antibodies of the present invention or antibodies derived from other species of the present invention can be produced by display-type techniques using techniques well known in the art, including but not limited to, phage display, retroviral display, ribosome display, and other techniques, the resulting molecules can be used for additional maturation such as affinity maturation, such techniques are well known in the art (see, for example, Hoogenboom et al., J.MoI.Biol.227.381 (1991) (phage display), Vaughan et al., Nature Biotech 14.309 (1996) (phage display), Hanes and Plucthau, PNASUSA 94, 4937-4942 (1997) (ribosome display), Parmley and Smith, Gene 73, 305-318 (1988) (phage display ), Scott TIBS 17, 241-245 (1992), Cwirla et al., PNAS USA 87, 6378-6382 (1990), Russet et al., Nucl. Acids Research 21, 1081-1085 (1993), Hoogenboom et al. , Immunol. Reviews 130, 43-68 (1992), Chiswell and McCafferty TIBTECH 10, 80-84 (1992), and US 5,733,743). If display techniques are used to generate non-human antibodies, such antibodies can be humanized, eg, as described elsewhere herein.

通过将人抗体的恒定结构域与非人物种的可变结构域相融合的方式可产生本发明的人源化单克隆抗体。如何制备人源化单克隆抗体的实施例可在例如US 6,054,297、US5,886,152和US 5,877,293中找到。人源化抗体被设计为与人免疫球蛋白比动物来源的单克隆抗体具有更高的同源性。来自“输入”(动物的)可变结构域的非人氨基酸残基典型地被转染到人“骨架”中。实质上，人源化可按照Winter及其同事(Jones et al.，Nature 321，522-525(1986)、Riechmann et al.，Nature 332，323-327(1988)、Verhoeyen et al.，Science 239，1534-1536(1988))的方法，通过将鼠补体决定区(“CDRs”)或CDR序列替换为人抗体的相应序列的方式来进行。因此，在这种“人源化”抗体中，人可变结构域的CDR部分已被非人物种的相应部分替代。这样，人源化抗体是典型的人抗体，其中某些CDR残基和某些可能的框架残基被啮齿类抗体的相似部位所替代。用于制备人源化抗体的人重链和轻链可变结构域的选择对降低抗原性至关重要。根据所谓的“最适”方法，筛选针对已知的人可变结构域序列完整文库的啮齿类抗体可变结构域的序列。然后，与啮齿类最接近的人序列作为人源化抗体的人框架(FR)(Sims et al.，J.Immunol.，151，2296(1993)、Chothia etal.，J.MoI.Biol.196，901(1987))。另一种方法使用了来源于特定轻链或重链亚组中所有人抗体的共有序列的特定框架。相同的框架可用于几个不同的人源化抗体(Carter etal.，PNAS USA 89，4285(1992)、Presta et al.，J.Immunol.151，2623(1993))。Humanized monoclonal antibodies of the invention can be produced by fusing constant domains of human antibodies to variable domains from a non-human species. Examples of how to prepare humanized monoclonal antibodies can be found, for example, in US 6,054,297, US 5,886,152 and US 5,877,293. Humanized antibodies are designed to have higher homology to human immunoglobulins than monoclonal antibodies of animal origin. Non-human amino acid residues from an "input" (animal's) variable domain are typically transfected into a human "backbone". In essence, humanization can be performed according to Winter and colleagues (Jones et al., Nature 321, 522-525 (1986), Riechmann et al., Nature 332, 323-327 (1988), Verhoeyen et al., Science 239 , 1534-1536 (1988)) by replacing the murine complement determining regions ("CDRs") or CDR sequences with the corresponding sequences of human antibodies. Thus, in such "humanized" antibodies, portions of the CDRs of human variable domains have been replaced by corresponding portions from a non-human species. Thus, humanized antibodies are typically human antibodies in which some CDR residues and possibly some framework residues are substituted by analogous sites from rodent antibodies. The choice of human heavy and light chain variable domains used to make humanized antibodies is critical to reducing antigenicity. According to the so-called "fittest" method, the sequences of the variable domains of rodent antibodies are screened against the complete library of known human variable domain sequences. Then, the human sequence closest to the rodent was used as the human framework (FR) of the humanized antibody (Sims et al., J.Immunol., 151, 2296 (1993), Chothia et al., J.MoI.Biol.196 , 901 (1987)). Another method uses a specific framework derived from the consensus sequence of all human antibodies in a specific subgroup of light or heavy chains. The same framework can be used for several different humanized antibodies (Carter et al., PNAS USA 89, 4285 (1992), Presta et al., J. Immunol. 151, 2623 (1993)).

典型地，人源化抗体保持对抗原高亲和性及其它有利的生物学特性也很重要。为了达到这个目的，可使用母序列和人源化序列的三维模型通过分析母序列及多种与之有关的人源化产物的过程来制备人源化抗体。通常三维免疫球蛋白模型是可获得，本领域技术人员对此比较了解。已有可示例并展现所选的候选免疫球蛋白序列的可能的三维构象结构的计算机程序。考察这些展示的结构可分析特定残基在候选免疫球蛋白序列的功能中的可能作用，即，对能影响候选免疫球蛋白与其抗原结合的能力进行分析。尽管CDR残基直接并基本上影响抗原结合，但通过这种方式，可从受体选择并组合FR残基，并输入序列，这样可使所需的诸如增加对靶抗原亲和性这样的抗体特征最大化。Typically, it is also important for humanized antibodies to retain high affinity for the antigen and other favorable biological properties. To this end, humanized antibodies can be prepared by a process of analysis of the parental sequence and various humanized products related thereto, using three-dimensional models of the parental and humanized sequences. Typically three-dimensional immunoglobulin models are available and are well understood by those skilled in the art. Computer programs are available which illustrate and display probable three-dimensional conformational structures of selected candidate immunoglobulin sequences. Examination of these displayed structures allows analysis of the likely role of particular residues in the function of the candidate immunoglobulin sequence, ie, the ability to affect the ability of the candidate immunoglobulin to bind its antigen. Although the CDR residues directly and substantially affect antigen binding, in this way FR residues can be selected and combined from the receptor and imported into sequence such that desired antibodies such as increased affinity for the target antigen can be achieved. feature maximization.

可使用任何合适的技术对鼠抗体或来自其它物种的抗体使其人源化或灵长类化，多种合适的技术已是本领域周知的(参见，例如Winterand Harris Immunol Today 14，43-46(1993)and Wright et al.，Crit.Reviewsin Immunol.125-168(1992))。目的抗体可通过重组DNA技术进行加工，以用相应的人序列来替代C_H1、C_H2、C_H3、铰链结构域和/或框架结构域(参见WO 92/02190和US 5,530,101、US 5,585,089、US5,693,761、US 5,693,792、US5,714,350和US 5,777,085)。Murine antibodies or antibodies from other species can be humanized or primatized using any suitable technique, many of which are well known in the art (see, e.g., Winter and Harris Immunol Today 14, 43-46 (1993) and Wright et al., Crit. Reviews in Immunol. 125-168 (1992)). The antibody of interest can be processed by recombinant DNA techniques to replace the _{CH1, CH2, CH3 , hinge} and/or framework domains with corresponding human sequences (see WO 92/02190 and US 5,530,101, US 5,585,089, US 5,693,761, US 5,693,792, US 5,714,350 and US 5,777,085).

也可按照Winter及其同事的方法(Jones et al.，Nature 321，522-525(1986)、Riechmann et al.，Nature 332，323-327(1988)、Verhoeyenet al.，Science 239，1534-1536(1988))，通过将啮齿类CDRs和CDR序列替换为相应的人抗体序列的方式来对抗体进行人源化。因此，这种“人源化”抗体在某种意义上是嵌合抗体(US 4,816,567)，其中实质上少于完整的人可变区结构域已被非人物种的相应序列所替代。实际上，典型地，人源化抗体是人抗体，其中某些CDR残基及某些可能的FR残基被来自啮齿类抗体中同源位点的残基所替代。Also according to the method of Winter and his colleagues (Jones et al., Nature 321, 522-525 (1986), Riechmann et al., Nature 332, 323-327 (1988), Verhoeyen et al., Science 239, 1534-1536 (1988)), humanizing antibodies by replacing rodent CDRs and CDR sequences with the corresponding human antibody sequences. Thus, such "humanized" antibodies are chimeric antibodies in the sense (US 4,816,567) in which substantially less than an intact human variable region domain has been replaced by the corresponding sequence from a non-human species. In fact, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted by residues from homologous sites in rodent antibodies.

此外，用于构建嵌合免疫球蛋白基因的Ig cDNA的用途在本领域是已知的(参见，例如Liu et al.，PNAS USA 84，3439(1987)and J.Immunol.139，3521(1987))。从杂交瘤或其它产生抗体的细胞中分离mRNA，用于产生cDNA：目的cDNA可通过使用特异引物的聚合酶链反应来扩增(US 4,683,195和US 4,683,202)。选择性地，制备并筛选文库，以分离目的序列。然后将编码可变区的DNA序列与人恒定区序列融合。人恒定区(及可变区)序列可在Kabat et al.，(1991)中找到，最近的相关数据可在http://www.biochem.ucl.ac.uk/～ martin/abs/Generallnfo.html获得。典型地，通过诸如补体结合或抗体依赖的细胞毒素作用活性这样的所需的效应物功能来指导同型体的选择。示例的同型体有IgG1、IgG2、IgG3和IgG4。可使用人轻链恒定区κ或λ的其中之一。然后可通过常规方法来表达嵌合的人源化抗体。Furthermore, the use of Ig cDNA for constructing chimeric immunoglobulin genes is known in the art (see, for example, Liu et al., PNAS USA 84, 3439 (1987) and J. Immunol. 139, 3521 (1987) )). Isolation of mRNA from hybridomas or other antibody-producing cells is used to generate cDNA: the cDNA of interest can be amplified by polymerase chain reaction using specific primers (US 4,683,195 and US 4,683,202). Optionally, libraries are prepared and screened to isolate sequences of interest. The DNA sequences encoding the variable regions are then fused to human constant region sequences. Human constant region (and variable region) sequences can be found in Kabat et al., (1991), and the most recent relevant data are available at http://www.biochem.ucl.ac.uk/~martin/abs/Generallnfo . html get. Isotype selection is typically guided by desired effector functions such as complement fixation or antibody-dependent cytotoxic activity. Exemplary isotypes are IgGl, IgG2, IgG3 and IgG4. One of the human light chain constant regions, kappa or lambda, can be used. The chimeric humanized antibodies can then be expressed by conventional methods.

就多聚体而言，本发明的CD38BPs可以是任何合适的形式。抗CD38抗体及抗体片段如果不是更高的多聚体形式，例如与IgM抗体相联的抗体，则至少是异源三聚体。在其它技术方案中，CD38BP可以二聚体或单体形式存在。例如，本发明的单体CD38BPs可被任何合适的技术所修饰，以形成多聚体肽组分。In terms of multimers, the CD38BPs of the invention may be in any suitable form. Anti-CD38 antibodies and antibody fragments are at least heterotrimeric, if not higher multimeric forms, such as antibodies linked to IgM antibodies. In other technical schemes, CD38BP can exist in the form of dimers or monomers. For example, monomeric CD38BPs of the invention can be modified by any suitable technique to form multimeric peptide components.

如果需要，本发明的抗CD38抗体的种类可通过已知方法进行转换。例如，最初为IgM的本发明的抗体可经类别转换为本发明的IgG抗体。此外，类别转换技术可用于将一种IgG亚类转换为另一种，例如，从IgG1转换为IgG2。因此，本发明中抗体的效应物功能可通过同型体转换改变为，例如，用于多种治疗用途的IgG1、IgG2、IgG3、IgG4、IgD、IgA、IgE或IgM抗体。The type of anti-CD38 antibody of the present invention can be switched by a known method, if necessary. For example, an antibody of the invention that is originally IgM can be class switched to an IgG antibody of the invention. In addition, class switching techniques can be used to switch one IgG subclass to another, for example, from IgG1 to IgG2. Thus, the effector functions of the antibodies of the invention can be altered by isotype switching to, for example, IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE or IgM antibodies for various therapeutic uses.

在一个技术方案中，本发明的抗体是IgG1抗体，例如IgG1，κ或 IgG1，λ同型体。在另一个技术方案中，本发明的抗体是IgG3抗体，例如IgG3，κ或IgG3，λ同型体。在另一个技术方案中，本发明的抗体是IgG4抗体，例如IgG4，κ或IgG4，λ同型体。在另一个技术方案中，本发明的抗体是IgA1或1gA2抗体。在另一个技术方案中，本发明的抗体是IgM抗体。In one technical solution, the antibody of the invention is an IgG1 antibody, such as an IgG1, kappa or IgG1, lambda isotype. In another embodiment, the antibody of the invention is an IgG3 antibody, such as IgG3, kappa or IgG3, lambda isotype. In another embodiment, the antibody of the invention is an IgG4 antibody, such as IgG4, kappa or IgG4, lambda isotype. In another technical solution, the antibody of the invention is an IgA1 or IgA2 antibody. In another technical solution, the antibody of the invention is an IgM antibody.

可从诸如scFv噬菌体展示文库这样的重组组合抗体文库中获得抗CD38抗体，它是用来源于人淋巴细胞的mRNA制备的人V_L和V_H cDNAs制成的。制备和筛选这类文库的方法在本领域是已知的。已有多种商品化的试剂盒用于生成噬菌体展示文库。还有其它的用于生成并筛选抗体展示文库的方法和试剂(参见，例如US 5,223,409、WO 92/18619、WO 91/17271、WO 92/20791、WO 92/15679、WO 93/01288、WO92/01047、WO 92/09690，Fuchs etal.，Bio/Technology 9，1370-1372(1991)，Hay et al.，Hum.Antibod.Hybridomas 3，81-85(1992)、Huseet al.，Science 246，1275-1281(1989)，McCafferty et al.，Nature 348，552-554(1990)、Griffiths et al.，EMBO J 12，725-734(1993)，Hawkinset al.，J.MoI.Biol.226.889-896(1992)、Clackson et al.，Nature 352，624-628(1991)，Gram etal.，PNAS USA 89，3576-3580(1992)，Garradet al.，Bio/Technology 9，1373-1377(1991)，Hoogenboom et ai.，Nuc AcidRes 19，4133-4137(1991)和Barbas et al.，PNASUSA 88，7978-7982(1991))。可使用任何适当的方法来筛选合适的V_L和V_H核酸序列。例如，可通过WO 93/06213中所述的抗原决定簇印迹方法来筛选V_L和V_H核酸。可使用诸如在例如WO92/01047，McCafferty et al.，Nature 348，552-554(1990)和Griffiths et al.，EMBOJ12，725-734(1993)所述的已知合适的方法来制备和筛选诸如scFv文库这样的抗体文库(以含人CD38的肽作为抗原)。这种抗体文库和CD38BPs的其它组合(文库、基因库等)是本发明的特征，它可用于医疗用途，以提供更全面的免疫应答；在免疫原性的肽、小分子、其它抗CD38抗体(例如，通过竞争检测)等的筛选方法中；和/或在诊断方法及组分(例如，通过标准技术可制备含可与其它抗体连接的这种抗体的点阵的免疫检测芯片)中作为工具。当选择初始的人V_L和V_H片段后，可进行“混合及匹配”实验，其中筛选与含CD38肽结合的初始选择的V_L和V_H片段的不同配对，以选取所需的V_L/V_H配对组合。例如，肽的反应性可通过ELISA或其它合 适的抗原决定簇分析方法来确定(参见，例如Scott，J.K.and Smith，G.P.Science249，386-390(1990)、Cwirla et al.，PNAS USA 87，6378-6382(1990)、Felici et al.，J.MoI.Biol.222，301-310(1991)and Kuwabaraet al.，Nature Biotechnology 15，74-78(1997)，以上文献对这些技术和原理进行讨论)。可通过抗体对抗原的亲和性和/或其从抗原解离的动力学(解离速率)来选择抗体(参见，例如Hawkins et al.，J.MoI.Biol.226，889-896(1992))。Anti-CD38 antibodies can be obtained from recombinant combinatorial antibody libraries such as scFv phage display libraries made from human _VL and _VH cDNAs prepared from mRNA derived from human lymphocytes. Methods for preparing and screening such libraries are known in the art. A variety of commercial kits are available for generating phage display libraries. There are other methods and reagents for generating and screening antibody display libraries (see, e.g., US 5,223,409, WO 92/18619, WO 91/17271, WO 92/20791, WO 92/15679, WO 93/01288, WO 92/ 01047, WO 92/09690, Fuchs et al., Bio/Technology 9, 1370-1372 (1991), Hay et al., Hum. Antibod. Hybridomas 3, 81-85 (1992), Huse et al., Science 246, 1275 -1281 (1989), McCafferty et al., Nature 348, 552-554 (1990), Griffiths et al., EMBO J 12, 725-734 (1993), Hawkins et al., J.MoI.Biol.226.889-896 (1992), Clackson et al., Nature 352, 624-628 (1991), Gram et al., PNAS USA 89, 3576-3580 (1992), Garrad et al., Bio/Technology 9, 1373-1377 (1991), Hoogenboom et ai., Nuc Acid Res 19, 4133-4137 (1991) and Barbas et al., PNASUSA 88, 7978-7982 (1991)). Suitable _VL and _VH nucleic acid sequences can be screened using any suitable method. For example, _VL and _VH nucleic acids can be screened by the epitope imprinting method described in WO 93/06213. Known suitable methods such as those described in, for example, WO92/01047, McCafferty et al., Nature 348, 552-554 (1990) and Griffiths et al., EMBOJ12, 725-734 (1993) can be used to prepare and screen compounds such as An antibody library (using a peptide containing human CD38 as an antigen) such as a scFv library. Such antibody libraries and other combinations of CD38BPs (libraries, gene banks, etc.) are a feature of the present invention, which can be used in medical applications to provide a more comprehensive immune response; in immunogenic peptides, small molecules, other anti-CD38 antibodies (e.g., by competition assays), etc.; and/or in diagnostic methods and components (e.g., immunoassay chips containing dotted arrays of such antibodies that can be linked to other antibodies can be prepared by standard techniques) as tool. Once the initial human _VL and _VH segments are selected, "mix and match" experiments can be performed in which different pairs of the initially selected _VL and _VH segments that bind to the CD38-containing peptide are screened to select the desired _VL /V _H pairing combination. For example, the reactivity of the peptides can be determined by ELISA or other suitable epitope analysis methods (see, for example, Scott, JK and Smith, GPScience 249, 386-390 (1990), Cwirla et al., PNAS USA 87, 6378-6382 (1990), Felici et al., J. MoI. Biol. 222, 301-310 (1991) and Kuwabara et al., Nature Biotechnology 15, 74-78 (1997), which discuss these techniques and principles). Antibodies can be selected by their affinity for the antigen and/or their kinetics of dissociation from the antigen (off-rate) (see, e.g., Hawkins et al., J. MoI. Biol. 226, 889-896 (1992 )).

为了进一步改善抗CD38抗体的质量和/或多样性，V_L/V_H对的V_L 及V_H片段可在例如V_H和/或V_L的CDR3区域中，以与天然免疫应答中负责抗体亲和力成熟的体内体细胞突变过程相似的过程进行随机突变。该体外亲和力成熟可通过用分别与V_H CDR3或V_L CDR3互补的PCR引物扩增V_H和V_L区域来实现，其中典型地引物在特定位点上“引入”四种核苷酸碱基随机混合物，这样获得的PCR产物编码在V_H和/或V_L CDR3区域中引入随机突变的V_H和V_L片段。可再次筛选与含CD38的肽结合的随机突变的V_H和V_L片段。In order to further improve the quality and/or diversity of anti-CD38 antibodies, the _VL and _VH fragments of the _VL / _VH pair can be located, for example, in the CDR3 region of _VH and/or _VL , in order to interact with those responsible for the antibody in the innate immune response. Affinity maturation is a process similar to the in vivo somatic mutation process for random mutations. This in vitro affinity maturation can be achieved by amplifying the _VH and _VL regions with PCR primers complementary to the _VH CDR3 or _VL CDR3, respectively, where typically the primers "introduce" four nucleotide bases at specific sites A random mixture, the PCR products thus obtained encode _VH and _VL segments with random mutations introduced in the _VH and/or _VL CDR3 regions. Randomly mutated _VH and _VL fragments can be rescreened for binding to CD38-containing peptides.

筛选后，可从展示包装体(例如，从噬菌体基因组中)中回收编码所选抗体的核酸，并通过标准重组DNA技术亚克隆到合适的载体中。如果需要，这种编码抗体的核酸可进行进一步操作，以生成其它抗体形式和CD38BPs。为了表达通过筛选组合文库分离的重组抗体，典型地，将含编码抗体序列的核酸克隆到重组表达载体中，并在适于表达核酸并产生抗体的条件下，转到合适的宿主细胞(哺乳动物细胞、酵母细胞等)内。Following selection, nucleic acid encoding the selected antibody can be recovered from the displayed package (eg, from the phage genome) and subcloned into a suitable vector by standard recombinant DNA techniques. Such antibody-encoding nucleic acids can be further manipulated to generate other antibody formats and CD38BPs, if desired. In order to express a recombinant antibody isolated by screening a combinatorial library, typically, the nucleic acid containing the sequence encoding the antibody is cloned into a recombinant expression vector, and transformed into a suitable host cell (mammalian) under conditions suitable for expressing the nucleic acid and producing the antibody. cells, yeast cells, etc.).

诸如人单链Fv(scFv)及Fab抗体片段这样的高亲和抗体肽也可使用将目标抗原固定在诸如微滴定板或珠这样的固体表面上的淘选技术从这类文库中来分离(参见，例如Barbas and Burton，Trends.Biotechnol.14，230-234(1996)and Aujame et al.，Hum.Antibodies 8，155-68(1997))。大型天然文库的噬菌体展示也使得不需免疫而直接分离人抗体成为可能(参见例如，de Haard et al.，J.Biol.Chem.274(26)，18218-18230(1999))。High affinity antibody peptides such as human single chain Fv (scFv) and Fab antibody fragments can also be isolated from such libraries using panning techniques that immobilize the antigen of interest on a solid surface such as microtiter plates or beads ( See, eg, Barbas and Burton, Trends. Biotechnol. 14, 230-234 (1996) and Aujame et al., Hum. Antibodies 8, 155-68 (1997)). Phage display of large natural libraries also enables the direct isolation of human antibodies without immunization (see eg, de Haard et al., J. Biol. Chem. 274(26), 18218-18230 (1999)).

在一个技术方案中，本发明提供变异的CD38抗体。“变异的”抗CD38抗体是与母抗体(典型地，通过免疫反应来产生)在CDRs或其它V_H和/或V_L序列中有一个或多个氨基酸改变，及替代、缺失、插入或 添加末端序列的抗体(即使这种改变没有改善母抗体抗原决定簇结合特性，但只要保留母抗体抗原决定簇结合特性最小实质量即可)。In one technical solution, the present invention provides a variant CD38 antibody. "Variant" anti-CD38 antibodies have one or more amino acid changes, substitutions, deletions, insertions or additions in the CDRs or other _VH and/or _VL sequences from the parent antibody (typically, produced by immunization) Antibodies with terminal sequences (even if such alterations do not improve the epitope binding properties of the parent antibody, so long as a minimal substantial amount of the epitope binding properties of the parent antibody are retained).

在单个变异抗体中的框架区、恒定结构域和/或可变区(或其任何一个或多个CDRs)的每个中可产生抗体变异体的变异形式。选择性地，可仅在抗体框架区、可变区(或其单个CDR)、或恒定结构域中的某一个产生变异。诸如由Cunningham and Wells，Science244，1081-1085(1989)所述的这样的丙氨酸扫描突变技术可用于在生成的含变异V_L、V_H或替代CDR序列的CD38BPs中鉴定用于替代或缺失的合适的残基，不过也可使用其它合适的突变技术。也可使用诸如Reidhaar-Olson andSauer，Science 241，53-57(1988)or Bowie andSauer，PNAS USA 86，2152-2156(1989)所述的已知突变和筛选方法来产生并检测单个氨基酸替代。Variant forms of antibody variants can be produced in each of the framework regions, constant domains and/or variable regions (or any one or more CDRs thereof) in a single variant antibody. Alternatively, variations can be made in only one of the antibody framework regions, variable regions (or individual CDRs thereof), or constant domains. Alanine scanning mutagenesis techniques such as those described by Cunningham and Wells, Science 244, 1081-1085 (1989) can be used to identify potential for substitutions or deletions in generated CD38BPs containing variant _VL , _VH or alternative CDR sequences suitable residues, although other suitable mutation techniques may also be used. Single amino acid substitutions may also be generated and tested using known mutagenesis and screening methods such as those described by Reidhaar-Olson and Sauer, Science 241, 53-57 (1988) or Bowie and Sauer, PNAS USA 86, 2152-2156 (1989).

因此，例如，在抗体变异体中，可将一个或单个氨基酸残基引入或插入到，或紧邻母抗体一个或多个高变区，譬如在一个或多个CDRs中。抗CD38抗体变异体可含有任意数目的插入的氨基酸残基，只要仍保留母抗体中抗原决定簇结合特征的最少实质量即可。举例来说，本发明的抗CD38抗体变异体可含有约1-30个插入的氨基酸残基，例如约1-10个、譬如例如约2-10个、例如2-5个或譬如约1-5个插入的氨基酸残基。同样，举例来说，本发明的抗CD38抗体变异体可含有约1-30个缺失的氨基酸残基，例如约1-10个、譬如例如约2-10个、例如2-5个或譬如约1-5个缺失的氨基酸残基。同样，举例来说，本发明的抗CD38抗体变异体可含有约1-30个替代的氨基酸残基，例如约1-10个、譬如例如约2-10个、例如2-5个或譬如约1-5个替代的氨基酸残基。同样，举例来说，本发明的抗CD38抗体变异体可含有约1-30个末端序列氨基酸残基增加，例如约1-10个、譬如例如约2-10个、例如2-5个或譬如约1-5个末端序列氨基酸残基增加。本发明的抗体变异体也包含2个或多个这种插入、缺失、替代及末端序列氨基酸残基增加的组合，只要这些变异体具有母抗体相对于一个或多个CD38抗原决定簇的亲和性、特异性和/或选择性的最小实质部分即可。Thus, for example, in an antibody variant, one or single amino acid residues may be introduced or inserted into, or immediately adjacent to, one or more hypervariable regions of the parent antibody, such as in one or more CDRs. Anti-CD38 antibody variants may contain any number of inserted amino acid residues so long as a minimum substantial amount of the epitope-binding characteristics of the parent antibody is retained. For example, an anti-CD38 antibody variant of the invention may contain about 1-30 inserted amino acid residues, such as about 1-10, such as for example about 2-10, such as 2-5 or such as about 1-10 5 inserted amino acid residues. Also, for example, an anti-CD38 antibody variant of the invention may contain about 1-30 deleted amino acid residues, such as about 1-10, such as about 2-10, such as 2-5 or such as about 1-5 missing amino acid residues. Also, for example, an anti-CD38 antibody variant of the invention may contain about 1-30 substituted amino acid residues, such as about 1-10, such as about 2-10, such as 2-5 or such as about 1-5 substituted amino acid residues. Also, for example, an anti-CD38 antibody variant of the invention may contain about 1-30 terminal sequence amino acid residue additions, such as about 1-10, such as for example about 2-10, such as 2-5 or such as About 1-5 terminal sequence amino acid residues were added. The antibody variants of the present invention also comprise combinations of 2 or more such insertions, deletions, substitutions, and terminal sequence amino acid residue additions, as long as these variants have the affinity of the parent antibody for one or more CD38 epitopes A minimum substantial portion of specificity, specificity and/or selectivity is sufficient.

在本文其它地方描述了在选择抗体变异体方面的考虑(例如，氨基酸残基功能特性的保守性、基于亲水特性的氨基酸残基的保守性，和/或基于分子量/大小的氨基酸残基的保守性)。典型地，诸如保守替代变 异这样的氨基酸序列改变预计不会对母序列结构特性造成实质改变(例如，替代的氨基酸不应该破坏作为母序列功能特性的二级结构)。在例如Proteins，Structures and Molecular Principles(Creighton，Ed.，W.H.Freeman andCompany，New York(1984))，Introduction to ProteinStructure(C.Branden andJ.Tooze，eds.，Garland Publishing，New York，N.Y.(1991))and Thornton et at.，Nature 354，105(1991)中描述已知的多肽二级和三级结构的例子。其它关于设计和构建肽变异体的原则在例如Collinet et al.，J Biol Chem 2Z5(23)，17428-33(2000)中有所讨论。Considerations in selecting antibody variants (e.g., conservation of functional properties of amino acid residues, conservation of amino acid residues based on hydrophilic properties, and/or conservation of amino acid residues based on molecular weight/size) are described elsewhere herein. conservative). Typically, amino acid sequence changes such as conservative substitution variations are not expected to result in substantial changes in the structural properties of the parent sequence (e.g., the substituted amino acids should not disrupt secondary structure that is a functional property of the parent sequence). In e.g. Proteins, Structures and Molecular Principles (Creighton, Ed., W.H. Freeman and Company, New York (1984)), Introduction to Protein Structure (C. Branden and J. Tooze, eds., Garland Publishing, New York, N.Y. (1991)) Examples of known polypeptide secondary and tertiary structures are described in and Thornton et at., Nature 354, 105 (1991). Other principles for designing and constructing peptide variants are discussed eg in Collinet et al., J Biol Chem 2Z5(23), 17428-33 (2000).

抗体的氨基酸序列变异体可通过向抗体编码核酸中引入合适的核苷酸改变(例如通过定点突变)或通过化学肽合成方式来获得。这种变异体包括，例如，在此处示例的抗体氨基酸序列中残基的缺失和/或插入和/或替代和/或末端序列增加。可产生缺失、插入和替代的任何组合，以获得所需的变异体，只要变异体具有母抗体抗原决定簇结合特性的最小实质部分即可。相对于母抗体的氨基酸序列的改变也可通过譬如改变糖基化位点的数目或位置来改变变异抗体相对于母抗体的翻译后加工过程。Amino acid sequence variants of antibodies can be obtained by introducing appropriate nucleotide changes into the antibody-encoding nucleic acid (eg, by site-directed mutagenesis) or by chemical peptide synthesis. Such variants include, for example, deletions and/or insertions and/or substitutions and/or terminal sequence additions of residues in the antibody amino acid sequences exemplified herein. Any combination of deletions, insertions, and substitutions may be made to obtain the desired variant, so long as the variant possesses a minimal substantial portion of the epitope-binding properties of the parent antibody. Changes in the amino acid sequence relative to the parent antibody can also alter the post-translational processing of the variant antibody relative to the parent antibody by, for example, altering the number or location of glycosylation sites.

本发明的变异抗体可包含诸如在CDRs这样的高变区的改变。含有这类CDR变异体的CD38BPs的实施例在本文其它部分有所描述，并且如上所述，这类CD38BPs可以是抗体。Variant antibodies of the invention may comprise changes in hypervariable regions, such as in the CDRs. Examples of CD38BPs containing such CDR variants are described elsewhere herein, and as noted above, such CD38BPs may be antibodies.

本发明的变异抗体可包含位于高变区之外的框架(FR)改变，例如在Fc区域，该改变可与诸如改变抗体功能或药物动力学特性这样的有利特性相关。例如，框架区或恒定结构域中的替代或其它修饰(插入、缺失、末端序列增加或其任何组合)与变异抗体相对于母抗体半衰期的增加相关，或可用于改变变异抗体相对于母抗体的免疫原性，以提供与另一个分子共价或非共价结合的位点，或改变补体结合这样的特性，例如导致Clq结合与CDC或FcγR结合与抗体依赖的细胞毒素作用(ADCC)的降低或增加。例如，可在重链恒定区的234、235、236、237、297、318、320和322位中一个或多个氨基酸残基处进行替代，从而可导致与未修饰的抗体相比较，在保持与抗原结合的同时在效应物功能上发生改变，参见US 5,624,821和US 5,648,260。进一步文献有WO00/42072，它揭示了具有改变的Fc区的抗体增加了ADCC，WO 94/29351，揭示了在C_H2结构域N末端区域具有突变的抗体改变了抗体与FcRI结合的能力，从而降低了抗体与Clq结合的能力，这随后降低了抗体结合补体的能力。此外，Shields et al.，J.Biol.Chem.276，6591-6604(2001)描述了组合变异体，它可改善FcγRIII与例如T256A/S298A、S298A/E333A和S298A/E333A/K334A的结合。Variant antibodies of the invention may contain framework (FR) changes located outside of the hypervariable regions, for example in the Fc region, which changes may be associated with favorable properties such as altered antibody function or pharmacokinetic properties. For example, substitutions or other modifications (insertions, deletions, terminal sequence additions, or any combination thereof) in the framework regions or constant domains are associated with increased half-life of the variant antibody relative to the parent antibody, or can be used to alter the half-life of the variant antibody relative to the parent antibody. Immunogenicity, to provide a site for covalent or non-covalent binding to another molecule, or to alter the properties of complement fixation, such as resulting in a reduction in Clq binding and CDC or FcγR binding and antibody-dependent cytotoxicity (ADCC) or increase. For example, substitutions may be made at one or more of amino acid residues in positions 234, 235, 236, 237, 297, 318, 320, and 322 of the heavy chain constant region, which may result in an increase in retention compared to an unmodified antibody. Alteration in effector function concomitant with antigen binding, see US 5,624,821 and US 5,648,260. Further references are WO00/42072, which discloses that antibodies with altered Fc regions have increased ADCC, WO 94/29351, which discloses that antibodies with mutations in the N-terminal region of the _CH2 domain alter the ability of antibodies to bind to FcRI, This reduces the ability of the antibody to bind Clq, which in turn reduces the ability of the antibody to bind complement. Furthermore, Shields et al., J. Biol. Chem. 276, 6591-6604 (2001) describe combinatorial variants that improve the binding of FcγRIII to eg T256A/S298A, S298A/E333A and S298A/E333A/K334A.

抗体的体外半衰期也可通过改变Ig恒定结构域或Ig类似的恒定结构域的拯救受体抗原决定簇来改善，从而使该分子不含有完整的C_H2结构域或完整的Ig Fc区域，参见US6,121,022和US 6,194,551。可进一步通过在Fc区域引入突变，例如252位苏氨酸替代为亮氨酸、254位苏氨酸替代为丝氨酸、256位苏氨酸替代为苯丙氨酸来增加体外半衰期，参见US6,277,375。The in vitro half-life of antibodies can also be improved by altering the rescue receptor epitope of the Ig constant domain or an Ig-like constant domain so that the molecule does not contain a complete _CH2 domain or a complete Ig Fc region, see US6,121,022 and US6,194,551. The in vitro half-life can be increased by introducing mutations into the Fc region, such as replacing threonine at position 252 with leucine, threonine at position 254 with serine, and threonine at position 256 with phenylalanine, see US6,277,375 .

在一个技术方案中，本发明提供变异的抗CD38抗体，其中抗体中潜在的T细胞抗原决定簇通过常规设计被降低或删除。这样，例如，在一个技术方案中，本发明提供“去免疫化的”抗CD38抗体，其中潜在的T细胞抗原决定簇被删除。去免疫化的抗CD38抗体的设计和构建可通过任何合适的已知技术来实现(参见，例如WO9852976中关于制备去免疫化抗体的方法)，当将这种如本发明所述的CD38BPs(例如，抗CD38变异抗体)给药时，希望可消除或实质降低在人中的免疫原性。In one technical solution, the present invention provides a variant anti-CD38 antibody, wherein potential T cell epitopes in the antibody are reduced or deleted by conventional design. Thus, for example, in one embodiment, the invention provides "deimmunized" anti-CD38 antibodies in which potential T cell epitopes have been deleted. The design and construction of deimmunized anti-CD38 antibodies can be achieved by any suitable known technique (see, for example, WO9852976 on the method of preparing deimmunized antibodies), when such CD38BPs according to the present invention (such as , anti-CD38 variant antibody) administration, it is hoped that the immunogenicity in humans can be eliminated or substantially reduced.

其它框架突变包括可降低对蛋白酶水解的敏感性、降低对氧化的敏感性，和/或在相关变异抗体中赋予或改变其它生理化学或功能特性的序列改变。框架的氨基酸序列改变也可导致变异抗体中相对于母抗体糖基化模式发生改变。改变是指删除一个或多个母抗体中发现的碳水化合物部分，和/或增加一个或多个母抗体中不存在的糖基化位点。抗体的糖基化典型地是N连接或O连接的。N连接是指碳水化合物部分与天冬氨酸残基的侧链连接。三肽序列天冬氨酸-X-丝氨酸和天冬氨酸-X-苏氨酸是常见的碳水化合物部分与天冬氨酸侧链酶连接的识别序列，其中X是除脯氨酸外的任何氨基酸。因此，多肽中的这两种三肽序列的存在是潜在的糖基化位点。O连接糖基化是指诸如N乙酰半乳糖胺、半乳糖或木糖等糖与羟基氨基酸的连接，最常见的是丝氨酸或苏氨酸，但也可使用5-羟基脯氨酸或5-羟基赖氨酸。可通过改变氨基酸序列，使其含一个或多个上述三肽序列(对N连接糖基化位点而言)来增加抗体的糖基化位点。也可通过对原始抗体序列进行一个或多个丝氨酸或苏氨酸的增加、 或替代而进行改变(对O连接糖基化而言)。Other framework mutations include sequence changes that may reduce susceptibility to proteolysis, reduce susceptibility to oxidation, and/or confer or alter other physiochemical or functional properties in associated variant antibodies. Amino acid sequence changes in the framework can also result in altered glycosylation patterns in the variant antibody relative to the parent antibody. Alteration refers to deletion of one or more carbohydrate moieties found in the parent antibody, and/or addition of one or more glycosylation sites not present in the parent antibody. Glycosylation of antibodies is typically N-linked or O-linked. N-linked refers to the attachment of the carbohydrate moiety to the side chain of an aspartic acid residue. The tripeptide sequences aspartate-X-serine and aspartate-X-threonine are common recognition sequences for enzymatic attachment of carbohydrate moieties to aspartate side chains, where X is other than proline any amino acid. Therefore, the presence of these two tripeptide sequences in the polypeptide is a potential glycosylation site. O-linked glycosylation refers to the attachment of a sugar such as N-acetylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, but 5-hydroxyproline or 5-hydroxyproline can also be used Hydroxylysine. Glycosylation sites of an antibody may be increased by altering the amino acid sequence to contain one or more of the tripeptide sequences described above (for N-linked glycosylation sites). Alterations may also be made by the addition, or substitution, of one or more serine or threonine to the original antibody sequence (for O-linked glycosylation).

抗体也可在不加通常与和Fc 297位Asn连接的碳水化合物连接的海藻糖单位的转染瘤中表达，以增强Fc对FcγRIII的亲和性，从而导致在存在NK细胞时抗体的ADCC增强，参见Shield et al.，J.Biol.Chem.277.26733(2002)。其它基于海藻糖化来改变糖基化的方法如Kyowa的WO 00/61739中所述。另外，可改变糖基化以改变CDC。进一步的参考文献有WO99/54342和Umana et al.，Nat.Biotechnol.17，176(1999)，描述了构建的CHO细胞系表达GntIII，导致具有改变的糖型和改善的ADCC活性的单克隆抗体的表达。Antibodies can also be expressed in transfectomas without the addition of trehalose units, usually linked to carbohydrates linked to Asn 297 of Fc, to enhance the affinity of Fc for FcγRIII, resulting in enhanced ADCC of antibodies in the presence of NK cells , see Shield et al., J. Biol. Chem. 277.26733 (2002). Other methods for altering glycosylation based on fucosylation are described in WO 00/61739 by Kyowa. Additionally, glycosylation can be altered to alter CDC. Further references are WO99/54342 and Umana et al., Nat. Biotechnol. 17, 176 (1999), describing the construction of CHO cell lines expressing GntIII, resulting in monoclonal antibodies with altered glycoforms and improved ADCC activity expression.

其它可能的制备新型抗CD38抗体的合适技术包括CDR步移突变、抗体链替换、“简约突变”(Balint and Larrick，Gene 137，109-118(1993))，和其它亲和力成熟技术(参见，例如Wu et al.，PNAS USA95，6037-42(1998))。抗体库克隆技术也可用于产生变异抗体(参见，例如WO 96/33279)。Other possible suitable techniques for making novel anti-CD38 antibodies include CDR step mutation, antibody chain replacement, "parsimony mutation" (Balint and Larrick, Gene 137, 109-118 (1993)), and other affinity maturation techniques (see, e.g., Wu et al., PNAS USA 95, 6037-42 (1998)). Antibody library cloning techniques can also be used to generate variant antibodies (see, eg, WO 96/33279).

有多种已知可产生CDR变异体的技术，任何合适的技术或其组合可用于本发明文中来产生实施例中的抗体CDRs的CDR变异体。这种技术的实施例包括删除非必需残基，如Studnicka et al.，ProteinEngineering 7，805-814(1994)所述(也参见Soderlind etal.，Immunotechnology.4(3-4)，279-85(1999)，CDR步移突变和其它人工亲和力成熟技术(参见，例如Yang et al.，Journal of Molecular Biology254(3).392-403(1995))，CDR替换技术，其中CDRs典型地从不同组的选择性地含合成的寡核苷酸的基因模板中扩增，并扩增V_L、V_H 的恒定区和/或CDRs，然后将各种片段混合(以单链或双链形式)，并通过聚合酶链反应(PCR)组装，从而产生一组携带引入到母框架中的替换的CDR的基因产物编码的抗体片段，该基因产物是用可与插入限制性位点之外的位点退火的外侧引物扩增的，从而保证产生全长的产物，将其插入到选自的载体中，用于表达含变异CDR的蛋白。可通过变异体/模拟结构和母序列的结构的叠印，例如，通过比较NMR溶液结构来测定合适的结构。合理设计CDR序列变异体的有用方法如例如WO91/09967和WO 93/16184中所述。这些方法的其它实施例在本文其它部分提供。There are a variety of known techniques for generating CDR variants, and any suitable technique or combination thereof can be used in the context of the present invention to generate CDR variants of the antibody CDRs of the Examples. Examples of this technique include deletion of non-essential residues as described by Studnicka et al., Protein Engineering 7, 805-814 (1994) (see also Soderlind et al., Immunotechnology. 4(3-4), 279-85( 1999), CDR walking mutagenesis and other artificial affinity maturation techniques (see, for example, Yang et al., Journal of Molecular Biology 254(3).392-403 (1995)), CDR replacement techniques, where CDRs are typically selected from different groups of selectively amplify in gene templates containing synthetic oligonucleotides, and amplify the constant regions and/or CDRs of _VL , _VH , and then mix the various fragments (in single-stranded or double-stranded form), and Assembled by polymerase chain reaction (PCR), resulting in a set of antibody fragments encoded by the gene product carrying the substituted CDRs introduced into the parent framework in a manner that anneals to a site other than the insertion restriction site Amplified by the outer primers to ensure the production of full-length products, which are inserted into selected vectors for the expression of proteins containing variant CDRs. Overprinting of the structure of the variant/mock structure and the parent sequence, such as , suitable structures are determined by comparison of NMR solution structures. Useful methods for the rational design of CDR sequence variants are described, for example, in WO 91/09967 and WO 93/16184. Additional examples of these methods are provided elsewhere herein.

本发明也提供具有与CD38结合能力的本发明的抗体(包括变异抗 体)的片段(CD38结合片段)。因此，CD38BPs包括含小于天然抗体整个四聚体结构的类似抗体的分子。抗体片段可以是任何含有全长抗体一部分的肽，通常为抗原结合或其可变区部分(这包括例如含本发明抗体的CDRs的人源化抗体片段、其变异体，或其它可使抗原片段与本发明的抗体竞争结合CD38的其它CDRs)。在一个技术方案中，抗体片段是指主要由或仅由抗体分子的一部分组成的肽。在一个技术方案中，本发明提供至少含包括本发明的一个或多个V_HCDRs的重链可变结构域，选择性地也含具有本发明的一个或多个V_L CDRs的轻链可变结构域的一部分的抗体片段，其中重链可变结构域及选择性地轻链可变结构域选择性地与其它部分，例如免疫球蛋白恒定结构域融合。恒定结构域序列可加到重链和/或轻链序列中，形成具有部分长度重链和/或轻链的种类。可使用任何抗体同型体恒定区或其部分，包括IgG、IgM、IgA、IgD和IgE的恒定区来实现该目的。The present invention also provides fragments (CD38-binding fragments) of the antibodies (including variant antibodies) of the present invention that have the ability to bind to CD38. Thus, CD38BPs include antibody-like molecules that contain less than the entire tetrameric structure of native antibodies. Antibody fragments can be any peptide that contains a portion of a full-length antibody, typically an antigen-binding or variable region portion thereof (this includes, for example, humanized antibody fragments comprising the CDRs of an antibody of the invention, variants thereof, or other fragments that can render the antigen other CDRs that compete with the antibodies of the invention for binding to CD38). In one technical solution, an antibody fragment refers to a peptide consisting mainly or only of a part of an antibody molecule. In one technical solution, the present invention provides at least a heavy chain variable domain comprising one or more _VH CDRs of the present invention, optionally also containing a light chain variable domain having one or more _VL CDRs of the present invention. Antibody fragments that are part of the variable domains in which the variable domains of the heavy chain and optionally the variable domains of the light chain are selectively fused to other parts, such as immunoglobulin constant domains. Constant domain sequences may be added to the heavy and/or light chain sequences to form species with partial length heavy and/or light chains. The constant region of any antibody isotype, or portion thereof, including the constant regions of IgG, IgM, IgA, IgD, and IgE, may be used for this purpose.

CD38结合抗体片段的例子包括Fab、Fab′、F(ab′)₂和Fv片段。本发明的抗体片段也包括含CDR等的肽。在一个技术方案中，本发明提供含具有任何此处所述的重链CDRs的第一多肽链和具有任何此处所述的轻链CDRs的第二多肽链，其中两个多肽链通过一个或多个链内二硫键共价连接。在一个技术方案中，本发明提供具有这种特征的两链抗体片段，其中抗体片段选自Fab、Fab′、Fab′-SH、Fv和/或F(ab′)₂片段。可通过常规方法使抗体片段化，可使用上述针对整个抗体的相同方法来筛选片段。例如，可通过胃蛋白酶处理抗体来产生F(ab′)₂片段。得到的F(ab′)₂片段可通过处理除去二硫键，从而产生Fab1片段。可通过木瓜蛋白酶处理IgG抗体获得Fab片段；可通过胃蛋白酶消化IgG抗体获得Fab′片段。将Fab1通过如下所述巯基或二硫键连接可产生Fab1片段。Fab1片段是通过切割F(ab′)₂的铰链区的二硫键而获得的抗体片段。可通过用还原剂，例如二硫苏糖醇处理F(ab′)₂片段而获得Fab′片段。抗体片段肽也可通过在重组细胞中表达编码这种肽的核酸来产生(参见，例如Evans etah，J.Immunol.Meth.184，123-38(1995))。例如，编码F(ab′)₂片段的一部分的嵌和基因可包括编码C_H1结构域和H链铰链区的DNA序列，然后通过翻译终止密码子来产生这种截短的抗体片段分子。Examples of CD38 binding antibody fragments include Fab, Fab', F(ab') ₂ and Fv fragments. The antibody fragments of the present invention also include peptides containing CDRs and the like. In one embodiment, the present invention provides a first polypeptide chain having any of the heavy chain CDRs described herein and a second polypeptide chain having any of the light chain CDRs described herein, wherein the two polypeptide chains are passed through One or more intrachain disulfide bonds are covalently linked. In one technical solution, the present invention provides a two-chain antibody fragment having such characteristics, wherein the antibody fragment is selected from Fab, Fab', Fab'-SH, Fv and/or F(ab') ₂ fragments. Antibodies can be fragmented by conventional methods, and the fragments can be screened using the same methods described above for whole antibodies. For example, F(ab') ₂ fragments can be produced by pepsinizing the antibody. The resulting F(ab') ₂ fragment can be treated to remove the disulfide bond, thereby generating a Fab1 fragment. Fab fragments can be obtained by papain treatment of IgG antibodies; Fab' fragments can be obtained by pepsin digestion of IgG antibodies. Fab1 fragments can be generated by linking Fab1 via sulfhydryl or disulfide bonds as described below. The Fab1 fragment is an antibody fragment obtained by cleavage of the disulfide bond of the hinge region of F(ab') ₂ . Fab' fragments can be obtained by treating F(ab') ₂ fragments with a reducing agent, such as dithiothreitol. Antibody fragment peptides can also be produced by expressing nucleic acids encoding such peptides in recombinant cells (see, eg, Evans etah, J. Immunol. Meth. 184, 123-38 (1995)). For example, a chimera gene encoding a portion of an F(ab') ₂ fragment may include DNA sequences encoding the CH1 domain and the _H chain hinge region, followed by translational stop codons to generate such truncated antibody fragment molecules.

CD38BPs也包括单价抗体和单链抗体。单链抗体是其中重链和轻链 Fv区连接的肽。在一个技术方案中，本发明提供单链Fv(scFv)，其中本发明的抗CD38抗体的Fv中的重链和轻链通过柔性的肽连接物(典型地约10、12、15或更多氨基酸残基)连接在单个肽链上。产生这种抗体的方法如例如US 4,946,778，Pluckthun in The Pharmacology ofMonoclonalAntibodies，vol.113，Rosenburg and Moore eds.Springer-Verlag，New York，pp.269-315(1994)，Bird et al.，Science 242，423-426(1988)、Huston et al.，PNAS USA 85，5879-5883(1988)及McCaffertyet al.，Nature 348，552-554(1990)所述。如果只使用单个V_H和V_L，则单链抗体可以是单价的，如果使用2个V_H和V_L，则是二价的，如果使用多于两个V_H和V_L，则是多价的。CD38BPs also include monovalent antibodies and single chain antibodies. Single chain antibodies are peptides in which the Fv regions of the heavy and light chains are linked. In one technical solution, the present invention provides single-chain Fv (scFv), wherein the heavy chain and light chain in the Fv of the anti-CD38 antibody of the present invention are connected by a flexible peptide linker (typically about 10, 12, 15 or more amino acid residues) linked to a single peptide chain. Methods for producing such antibodies are described, for example, in US 4,946,778, Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269-315 (1994), Bird et al., Science 242, 423-426 (1988), Huston et al., PNAS USA 85, 5879-5883 (1988) and McCafferty et al., Nature 348, 552-554 (1990). ScFvs can be monovalent if only a single _VH and _VL is used, bivalent if 2 _VH and _VL are used, or polyvalent if more than two _VH and _VL are used price.

在本发明的一个技术方案中，CD38BP可来源于或与另一个功能分子，例如另一个肽或蛋白(例如Fab′片段)连接，从而产生与多个结合位点或靶抗原决定簇结合的双特异或多特异分子。例如，本发明的抗体可功能上与一个或多个其它结合分子，例如另一个抗体、肽或结合类似物连接(例如，通过化学偶联、基因融合、非共价连接或其它方式)。在一个技术方案中，CD38BP是本发明的抗体。In one technical solution of the present invention, CD38BP can be derived from or linked to another functional molecule, such as another peptide or protein (such as a Fab' fragment), thereby generating a double binding site or target antigenic determinant. Specific or multispecific molecules. For example, an antibody of the invention can be functionally linked (eg, by chemical conjugation, genetic fusion, non-covalent linkage or otherwise) to one or more other binding molecules, such as another antibody, peptide or binding analog. In one technical solution, CD38BP is the antibody of the present invention.

因此，本发明包括含至少一个对CD38的第一结合特异性和针对第二个靶抗原决定簇的第二结合特异性的双特异和多特异分子。在本发明的一个技术方案中，第二个靶抗原决定簇是Fc受体，例如人FcγRI(CD64)或人Fcα受体(CD89)，或T细胞受体，例如CD3。在一个技术方案中，本发明提供可与表达FoγR、FcαR或FcεR的效应细胞(例如，单核细胞、巨噬细胞或分叶核细胞(PMNs))结合，并作用于表达CD38的细胞的双特异和多特异分子。这些双特异和多特异分子使表达CD38的细胞定位到效应细胞，并启动诸如表达CD38细胞的吞噬作用、抗体依赖的细胞毒素作用(ADCC)、细胞因子释放、或过氧化物阴离子的产生这样的Fc受体介导的效应细胞活性。Thus, the invention includes bispecific and multispecific molecules comprising at least a first binding specificity for CD38 and a second binding specificity for a second target epitope. In one embodiment of the invention, the second target epitope is an Fc receptor, such as human FcγRI (CD64) or human Fcα receptor (CD89), or a T cell receptor, such as CD3. In one technical solution, the present invention provides a dual-targeted drug that can bind to effector cells expressing FoγR, FcαR or FcεR (for example, monocytes, macrophages or segmented nuclear cells (PMNs)) and act on cells expressing CD38. Specific and multispecific molecules. These bispecific and multispecific molecules localize CD38-expressing cells to effector cells and initiate events such as phagocytosis of CD38-expressing cells, antibody-dependent cytotoxicity (ADCC), cytokine release, or superoxide anion production. Fc receptor-mediated effector cell activity.

本发明的双特异和多特异分子除了抗Fc结合特异性和抗CD38结合特异性之外，还可进一步包括第三种结合特异性。在一个技术方案中，第三种结合特异性是抗增强因子(EF)部分，例如，与表面蛋白结合的分子参与细胞毒素作用，因此，增加对靶细胞的免疫反应。“抗增强因子部分”可以是与给定分子结合的抗体，功能性抗体片段或配体，例如，抗原或受体，因此导致结合决定簇对Fc受体或靶细胞抗原的增强作用。 “抗增强因子部分”可结合Fc受体或靶细胞抗原。选择性地，抗增强因子部分可与不同于可结合第一和第二结合特异性的实体相结合。例如，抗增强因子部分可结合细胞毒素T细胞(例如，通过CD2、CD3、CD8、CD28、CD4、CD40、ICAM-1或针对靶细胞具有更高免疫应答的其它免疫细胞)。The bispecific and multispecific molecules of the present invention may further include a third binding specificity in addition to the anti-Fc binding specificity and anti-CD38 binding specificity. In one embodiment, the third binding specificity is an anti-enhancement factor (EF) moiety, eg, a molecule that binds to a surface protein involved in cytotoxicity, thereby increasing the immune response to target cells. An "anti-enhancer moiety" may be an antibody, a functional antibody fragment or a ligand that binds to a given molecule, eg, an antigen or a receptor, thus resulting in an enhanced effect of the binding determinant on an Fc receptor or target cell antigen. An "anti-enhancement factor portion" can bind an Fc receptor or a target cell antigen. Alternatively, the anti-enhancement factor portion can bind to an entity different from that to which the first and second binding specificities can bind. For example, anti-enhancement factor moieties can bind cytotoxic T cells (eg, via CD2, CD3, CD8, CD28, CD4, CD40, ICAM-1 or other immune cells with higher immune responses against target cells).

在一个技术方案中，本发明的双特异和多特异分子包含至少一个具有结合特异性的其它抗体，这种抗体包括例如Fab、Fab1、F(ab′)₂、Fv或scFv。该其它抗体也可以是轻链或重链二聚体，或诸如Fv或Ladneret al在US 4,946,778中所述的单链构建体这样的任何最小片段。抗体还可以是如US 2003/0118592和US 2003/0133939所述的结合结构域免疫球蛋白融合蛋白。In one embodiment, the bispecific and multispecific molecules of the invention comprise at least one other antibody with binding specificity, such antibody including, for example, Fab, Fab1, F(ab') ₂ , Fv or scFv. The other antibodies may also be light or heavy chain dimers, or any minimal fragment such as Fv or single chain constructs as described by Ladner et al in US 4,946,778. The antibody may also be a binding domain immunoglobulin fusion protein as described in US 2003/0118592 and US 2003/0133939.

在一个技术方案中，对Fc受体的结合特异性是由人单克隆抗体来提供的，其结合不被人免疫球蛋白G(IgG)所阻断。此处所用术语“IgG受体”是指位于染色体1上的8个γ链基因中的任何一个。这些基因共编码12个可分属于3个Fc^*受体种类：FcγRI(CD64)、FcγRII(CD32)和FcγRIII(CD16)转膜或可溶性受体同型体。在一个技术方案中，Fcγ受体是人高亲和性FcγRI。Fanger等在WO 88/00052和US 4,954,617中描述了这些单克隆抗体的产生和特性。这些抗体在不同于受体的Fcγ结合位点处与FcγRI、FcγRII或FcγRIII的抗原决定簇结合，因此，它们的结合不会被生理水平的IgG完全阻断。本发明中所用的特异性抗FcγRI抗体有mAb 22、mAb 32、mAb 44、mAb 62和mAb 197。在其它技术方案中，抗Fcγ受体抗体是mAb22(H22)的人源化形式。H22抗体的产生和特性在Graziano，R.F.et al.，J.Immunol.155(10)，4996-5002(1995)和WO 94/10332中有所描述。产生H22抗体的细胞系于1992年11月4曰，以HA022CL1的命名保存在美国典型培养物保藏中心，其序列号为CRL11177。In one embodiment, binding specificity for Fc receptors is provided by human monoclonal antibodies, the binding of which is not blocked by human immunoglobulin G (IgG). The term "IgG receptor" as used herein refers to any one of the eight gamma chain genes located on chromosome 1. These genes encode a total of 12 transmembrane or soluble receptor isotypes that can be classified into three Fc ^* receptor classes: FcγRI (CD64), FcγRII (CD32) and FcγRIII (CD16). In one embodiment, the Fcγ receptor is human high affinity FcγRI. Fanger et al. describe the production and characterization of these monoclonal antibodies in WO 88/00052 and US 4,954,617. These antibodies bind to epitopes of FcγRI, FcγRII or FcγRIII at an Fcγ-binding site distinct from the receptor and, therefore, their binding is not completely blocked by physiological levels of IgG. The specific anti-FcγRI antibodies used in the present invention are mAb 22, mAb 32, mAb 44, mAb 62 and mAb 197. In other embodiments, the anti-Fcγ receptor antibody is a humanized form of mAb22 (H22). The production and characterization of the H22 antibody is described in Graziano, RF et al., J. Immunol. 155(10), 4996-5002 (1995) and WO 94/10332. The cell line producing the H22 antibody was deposited in the American Type Culture Collection under the designation HA022CL1 on November 4, 1992, and its serial number is CRL11177.

在一个技术方案中，Fc受体的结合特异性是由与人IgA受体，例如Fcα受体(Fcα1(CD89))相结合的抗体来提供的，在一个技术方案中，其结合不会被人免疫球蛋白A(IgA)所阻断。术语“IgA受体”是指含有位于染色体19上的一个α基因(FcαRI)的基因产物。已知该基因编码几个55到110kDa选择性拼接的转膜同型体。在单核细胞/巨噬细胞、嗜曙红细胞和嗜中性粒细胞的FcαRI(CD89)是组成型表达的，但在非 效应细胞群中不是这样。FcαRI对IgA1和IgA2具有中等亲和性，当与诸如G-CSF或GM-CSF这样的细胞因子接触后亲和性会提高(Morton，H.C.et al.，Critical Reviews in Immunology 16，423-440(1996))。鉴定为A3、A59、A62和A77的、可在IgA配体结合结构域之外结合FcαRI的4种FcαRI-特异性单克隆抗体已被描述(Monteiro，R.C.et al.，J.Immunol.148，1764(1992))。In one technical solution, the binding specificity of the Fc receptor is provided by an antibody that binds to a human IgA receptor, such as the Fcα receptor (Fcα1 (CD89)), and in one technical solution, its binding is not inhibited Blocked by human immunoglobulin A (IgA). The term "IgA receptor" refers to the gene product comprising an alpha gene (FcaRI) located on chromosome 19. This gene is known to encode several alternatively spliced transmembrane isoforms of 55 to 110 kDa. FcαRI (CD89) is constitutively expressed in monocytes/macrophages, eosinophils and neutrophils, but not in non-effector cell populations. FcαRI has moderate affinity for IgA1 and IgA2, which increases when exposed to cytokines such as G-CSF or GM-CSF (Morton, H.C. et al., Critical Reviews in Immunology 16, 423-440( 1996)). Four FcαRI-specific monoclonal antibodies, identified as A3, A59, A62 and A77, that bind FcαRI outside the ligand-binding domain of IgA have been described (Monteiro, R.C. et al., J. Immunol. 148, 1764 (1992)).

FcαRI、FcγRI、FcγRII和FcγRIII，特别是FcγRII和FcγRIII是本发明中所用的触发受体的实施例，这是因为它们(1)主要在免疫效应细胞，例如单核细胞、PMNs、巨噬细胞和枝状细胞上表达；(2)以高水平表达(例如，每个细胞5,000-100,000)；(3)是细胞毒素作用的介导物(例如ADCC、吞噬作用)；及(4)介导增强的抗原，包括自身抗原，针对它们的抗原呈递。FcαRI, FcγRI, FcγRII, and FcγRIII, particularly FcγRII and FcγRIII, are examples of triggering receptors used in the present invention because they (1) are primarily found in immune effector cells such as monocytes, PMNs, macrophages and clamocytes. (2) expressed at high levels (e.g., 5,000-100,000 per cell); (3) are mediators of cytotoxic effects (e.g., ADCC, phagocytosis); and (4) mediate enhanced Antigens, including self-antigens, are presented against their antigen.

在一个技术方案中，本发明的CD38BP是多特异性抗CD38抗体或类似抗体的分子，其特定的实施例是包含至少一对特异针对含有至少部分CD38的抗原决定簇的V_H序列和V_L序列链，及另一个至少一对特异针对第二个抗原决定簇的V_H和V_L序列链的双特异性抗体。双特异抗体中的V_H和V_L序列可包含对应于抗CD38 V_H和V_L区、变异体V_H和/或V_L序列、或V_H和/或V_L区的合适部分的完整V_H和V_L序列，譬如，CDR序列和足以提供与目的抗原决定簇结合的其它序列的合适组合。In one technical solution, the CD38BP of the present invention is a multispecific anti-CD38 antibody or antibody-like molecule, a specific embodiment of which comprises at least one pair of _VH sequences and _VL sequences specific for an antigenic determinant containing at least part of CD38. sequence chain, and another at least one pair of bispecific antibodies specific for the _VH and _VL sequence chains of the second antigenic determinant. The _VH and _VL sequences in the diabodies may comprise complete V sequences corresponding to anti-CD38 _VH and _VL regions, variant _VH and/or _VL sequences, or suitable portions of _VH and/or _VL regions. A suitable combination of _H and _VL sequences, eg, CDR sequences and other sequences sufficient to provide binding to the antigenic determinant of interest.

示例的双特异性抗体分子包括(i)两种相互连接的抗体，一种特异针对CD38，第二种针对第二个靶标，(ii)具有一条特异针对CD38的链和第二条特异针对第二个分子的链的单个抗体，及(iii)特异针对CD38和第二种分子的单链抗体。典型地，第二个靶/第二种分子是除CD38外的分子。在一个技术方案中，第二种分子是癌抗原/肿瘤相关抗原，譬如癌胚抗原(CEA)、前列腺特异抗原(PSA)、RAGE(肾抗原)、α-胎蛋白、CAMEL(在黑色瘤上识别CTL的抗原)、CT抗原(譬如MAGE-B5、-B6、-C2、-C3和D；Mage-12；CT10；NY-ESO-1、SSX-2、GAGE、BAGE、MAGE和SAGE)、粘液素抗原(例如MUC1、粘液素CA125等)、神经节苷脂抗原、酪氨酸酶、gp75、C-myc、Marti、MelanA、MUM-1、MUM-2、MUM-3、HLA-B7和Ep-CAM。在一个技术方案中，第二个分子是诸如α5β3整联蛋白这样的癌相关整联蛋白。在一个技术方案中，第二个分子是血管生长因子或其它癌相关生长因子，譬如血管 内皮生长因子(VEGF)、成纤维细胞生长因子(FGF)、表皮生长因子(EGF)、表皮生长因子受体(EGFR)、血管生长素及其受体，特别是与癌进展(例如HER1-HER4受体之一)相关的受体。其它此处所述的癌进展相关的蛋白也是合适的第二种分子。在一个技术方案中，第二种分子是在诸如CD138这样的多发性骨髓瘤细胞表面表达的分子。Exemplary bispecific antibody molecules include (i) two interconnected antibodies, one specific for CD38 and the second for a second target, (ii) having one chain specific for CD38 and the second chain specific for the second target A single antibody for chains of both molecules, and (iii) a single chain antibody specific for CD38 and the second molecule. Typically, the second target/second molecule is a molecule other than CD38. In one technical solution, the second molecule is a cancer antigen/tumor-associated antigen, such as carcinoembryonic antigen (CEA), prostate-specific antigen (PSA), RAGE (renal antigen), α-fetoprotein, CAMEL (on melanoma Antigens that recognize CTL), CT antigens (such as MAGE-B5, -B6, -C2, -C3, and D; Mage-12; CT10; NY-ESO-1, SSX-2, GAGE, BAGE, MAGE, and SAGE), Mucin antigens (such as MUC1, mucin CA125, etc.), ganglioside antigens, tyrosinase, gp75, C-myc, Marti, MelanA, MUM-1, MUM-2, MUM-3, HLA-B7 and Ep-CAM. In one embodiment, the second molecule is a cancer-associated integrin such as α5β3 integrin. In one embodiment, the second molecule is an angiogenic growth factor or other cancer-associated growth factor, such as vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), epidermal growth factor (EGF), epidermal growth factor receptor body (EGFR), angiogenin and its receptors, especially those associated with cancer progression (eg, one of the HER1-HER4 receptors). Other cancer progression-associated proteins described herein are also suitable second molecules. In one embodiment, the second molecule is a molecule expressed on the surface of multiple myeloma cells, such as CD138.

在一个技术方案中，本发明的双特异抗体是双功能抗体。In one technical solution, the bispecific antibody of the present invention is a bifunctional antibody.

双特异抗体也包括交联的或“异源连接物”抗体。例如，在异源连接物中的抗体可与针对生物素的亲和素和其它分子偶联。例如，已计划使用这种抗体使免疫系统细胞可针对另外的细胞(参见，例如US4,676,980)。可使用任何便利的交联方法来生成异源连接物抗体。合适的肽交联试剂和技术在本领域是周知的，例如在US 4,676,980中揭示了这类试剂和技术的实施例。Bispecific antibodies also include cross-linked or "heterolinker" antibodies. For example, antibodies in the heterolinker can be conjugated to avidin and other molecules against biotin. For example, it has been planned to use such antibodies to target immune system cells against additional cells (see eg US 4,676,980). Any convenient cross-linking method can be used to generate heterolinker antibodies. Suitable peptide crosslinking reagents and techniques are well known in the art, examples of such reagents and techniques are disclosed, for example, in US 4,676,980.

因此，尽管此处讨论的是抗体，但应理解，如果合适，抗体的细节和特征可等价用于诸如Fab片段、Fab1片段和scFv肽、类似抗体的肽(含CDR的肽)、双和多特异抗体和其它CD38BPs的抗体片段，只要本发明的CD38BP保留相应完整抗体的抗原结合特性的最小实质部分即可。在某些情况下，抗体片段可具有较低的抗原结合亲和性，但可提供能弥补亲和性上任何这种损失的其它优势特征。Thus, although antibodies are discussed herein, it is to be understood that, where appropriate, the details and characteristics of antibodies are equivalently applicable to peptides such as Fab fragments, Fab1 fragments and scFv peptides, antibody-like peptides (CDR-containing peptides), bis and Antibody fragments of multispecific antibodies and other CD38BPs as long as the CD38BPs of the invention retain a minimal substantial portion of the antigen-binding properties of the corresponding intact antibodies. In certain instances, antibody fragments may have lower antigen-binding affinity, but may offer other advantageous characteristics that compensate for any such loss in affinity.

本发明的CD38BPs，特别是抗CD38抗体可基于它们能或不能提供补体结合的能力来进行选择。存在多种能进行补体结合和CDC的抗体同型体，包括但不限定于以下：鼠IgM、鼠IgG2a、鼠IgG2b、鼠IgG3、人IgM、人IgG1和人IgG3。这些不包括，也不限定的同型体鼠人IgG2和人IgG4。同型体测定和其它改变抗体的补体结合和CDC功能特征的方法是本领域已知的。本发明的CD38BPs也包括免疫黏附素，它是指其中抗CD38抗体的一个或多个CDRs与该分子共价或非共价连接的分子。免疫黏附素可与CDR(s)整合作为更大的多肽链的一部分，可将CDR(s)与另一种多肽链共价连接，或可非共价整合到CDR(s)中。CDRs可使免疫黏附素与CD38特异连接。The CD38BPs of the invention, particularly anti-CD38 antibodies, can be selected based on their ability or inability to provide complement fixation. There are a variety of antibody isotypes capable of complement fixation and CDC, including but not limited to the following: murine IgM, murine IgG2a, murine IgG2b, murine IgG3, human IgM, human IgGl and human IgG3. These do not include, and are not limited to, the isotypes murine human IgG2 and human IgG4. Isotype determination and other methods of altering the complement fixation and CDC functional characteristics of antibodies are known in the art. CD38BPs of the present invention also include immunoadhesins, which refer to molecules in which one or more CDRs of an anti-CD38 antibody are covalently or non-covalently linked to the molecule. Immunoadhesins can be integrated with CDR(s) as part of a larger polypeptide chain, can covalently link the CDR(s) to another polypeptide chain, or can be non-covalently integrated into the CDR(s). CDRs can specifically link immunoadhesin to CD38.

本发明也提供CD38BP融合蛋白。CD38BP融合蛋白可包含任何合适的氨基酸序列或特异和/或选择性地针对至少部分包含在CD38中的至少一个结构域的序列的组合(例如，抗CD38抗体V_H结构域、V_L结构域或特别是其CDRs)，及至少一个非同源的，典型地实质不相同的氨基酸序列(例如，与CD38特异的/选择性的序列的氨基酸同一性低于约40％、低于约35％、低于约30％、低于约25％、低于约20％)，它可赋予融合蛋白那些无法单独由CD38特异/选择序列所提供的可检测的生物功能和/或特性(例如，体内半衰期的延长、荧光、更多地作用于对特定细胞类型等)。这类融合蛋白的功能序列可被柔性连接物分开。二级序列也可来源于细胞毒性或凋亡的肽。二级序列也可提供诊断特性。这类序列的实施例包括那些诸如辣根过氧化物酶这样的易于可视化的酶的序列。The invention also provides CD38BP fusion proteins. A CD38BP fusion protein may comprise any suitable amino acid sequence or combination of sequences specific and/or selective for at least one domain comprised at least in part in CD38 (e.g., an anti-CD38 antibody _VH domain, _VL domain or particularly its CDRs), and at least one non-homologous, typically substantially non-identical, amino acid sequence (e.g., less than about 40%, less than about 35%, less than about 35% amino acid identity to a CD38-specific/selective sequence, less than about 30%, less than about 25%, less than about 20%), it can impart detectable biological functions and/or properties (e.g., in vivo half-life) to the fusion protein that cannot be provided by CD38 specific/selection sequences alone prolongation, fluorescence, more specific cell types, etc.). The functional sequences of such fusion proteins can be separated by flexible linkers. Secondary sequences can also be derived from cytotoxic or apoptotic peptides. Secondary sequences can also provide diagnostic properties. Examples of such sequences include those of easily visualized enzymes such as horseradish peroxidase.

CD38BP融合蛋白也可通过比较比较抗原决定簇标签来描述。抗原决定簇标签序列是具有足够多残基、可提供抗原决定簇用于针对生成的抗体的氨基酸序列，但在CD38BP中，该序列很短，以至于无法实质上干扰CD38BP的活性(选择性、特异性、亲和性和/或生物活性)(与缺失抗原决定簇标签的母CD38BP相比较而言)。所需的抗原决定簇标签应充分特异，从而可使抗抗原决定簇标签抗体实质上不与其它抗原决定簇发生交叉反应。合适的标签多肽一般具有至少约6个氨基酸残基，通常为8-50个氨基酸残基(例如，约9-30残基)。抗原决定簇标签的实施例包括flu HA标签多肽及其抗体12CA5(Field et ah，MoI.Cell.Biol.8，2159-2165(1988))；myc标签及其抗体8F9、3C7、6E10、G4、B7和9E10(Evan et al.，MoI.Cell.Biol.5(12)，3610-3616(1985))，和单纯疱疹病毒糖蛋白D(gD)标签及其抗体(Paborsky et al.，ProteinEngineering 3(6)，547-553(1990))。在特定技术方案中，抗原决定簇标签是“拯救受体结合抗原决定簇”。此处所用术语“拯救受体结合抗原决定簇”是指IgG分子中Fc区的抗原决定簇(IgG1、IgG2、IgG3、或gG4)，它负责增加IgG分子在体内的血清半衰期。CD38BP fusion proteins can also be described by comparing epitope tags. An epitope tag sequence is an amino acid sequence of sufficient residues to provide an epitope for antibodies raised against, but in CD38BP, the sequence is so short that it does not substantially interfere with CD38BP activity (selectivity, specificity, affinity and/or biological activity) (compared to parent CD38BP lacking the epitope tag). The desired epitope tag should be sufficiently specific such that the anti-epitope tag antibody does not substantially cross-react with other epitopes. Suitable tag polypeptides generally have at least about 6 amino acid residues, usually 8-50 amino acid residues (eg, about 9-30 residues). Examples of epitope tags include flu HA tag polypeptide and its antibody 12CA5 (Field et ah, MoI. Cell. Biol. 8, 2159-2165 (1988)); myc tag and its antibodies 8F9, 3C7, 6E10, G4, B7 and 9E10 (Evan et al., MoI.Cell.Biol.5(12), 3610-3616(1985)), and herpes simplex virus glycoprotein D (gD) tag and its antibody (Paborsky et al., ProteinEngineering 3 (6), 547-553 (1990)). In a particular embodiment, the epitope tag is a "rescue receptor binding epitope". The term "rescue receptor binding epitope" as used herein refers to an epitope in the Fc region of an IgG molecule (IgG1, IgG2, IgG3, or IgG4) that is responsible for increasing the serum half-life of the IgG molecule in vivo.

本发明的CD38BPs也包括CD38BP衍生物。衍生物是肽，其中肽的一个或多个氨基酸残基被化学修饰(例如，被烷基化、酰化、形成酯或形成氨基)或与一个或多个异源替代物(例如，亲脂性替代，PEG基团、与合适的有机基团连接物相连接的肽侧链等)共价连。肽也可与诸如细胞毒素、化疗药物、免疫抑制剂或放射性同位素(即所谓的免疫连接物)这样的治疗基团相连接。通常，此处所述的CD38BPs可被含有任何合适数目的这种修饰氨基酸的物质所修饰，和/或与这种连接替代物相联。在本文中适用性通常是通过至少在实质上保留与非衍生的母 CD38BP所具有的CD38选择性和/或特异性的能力来确定的。含有一个或多个修饰氨基酸的优势在于，例如(a)增加多肽血清半衰期，(b)降低多肽抗原性，或(c)增加多肽保藏稳定性。例如，氨基酸可在重组生产过程中，在翻译时或翻译后被修饰(例如，在哺乳动物细胞中表达时，在N-X-S/T元件上的N连接糖基化)，或通过合成方式来修饰。修饰氨基酸的非限制性实施例包括糖基化氨基酸、硫酸盐氨基酸、prenlyated(例如，法尼基化、geranylgeranylated)氨基酸、乙酰化氨基酸、酰化氨基酸、PEG化氨基酸、生物素化氨基酸、羧基化氨基酸、磷酸化氨基酸等。指导技术人员进行氨基酸修饰的参考文献遍布各种文献中。实验流程可在Walker(1998)Protein Protocols On Cd-Rom，HumanaPress，Towata，NJ中找到。修饰氨基酸可选自糖基化氨基酸、PEG化氨基酸、法尼基化氨基酸、乙酰化氨基酸、生物素化氨基酸、与脂基团连接的氨基酸及与有机衍生试剂连接的氨基酸。CD38BPs of the present invention also include CD38BP derivatives. Derivatives are peptides in which one or more amino acid residues of the peptide are chemically modified (e.g., alkylated, acylated, ester-forming, or amino-forming) or with one or more heterologous substitutes (e.g., lipophilic Alternatively, a PEG group, a peptide side chain attached to a suitable organic group linker, etc.) is covalently linked. Peptides can also be linked to therapeutic groups such as cytotoxins, chemotherapeutic drugs, immunosuppressants or radioisotopes (so-called immunoconjugates). In general, the CD38BPs described herein may be modified by substances containing any suitable number of such modified amino acids, and/or associated with such linkage surrogates. Suitability in this context is generally determined by the ability to retain at least substantially the CD38 selectivity and/or specificity possessed by the non-derived parent CD38BP. The advantage of having one or more modified amino acids is, for example, to (a) increase the serum half-life of the polypeptide, (b) reduce the antigenicity of the polypeptide, or (c) increase the storage stability of the polypeptide. For example, amino acids can be modified during recombinant production, during translation or post-translationally (eg, N-linked glycosylation on N-X-S/T elements when expressed in mammalian cells), or synthetically. Non-limiting examples of modified amino acids include glycosylated amino acids, sulfated amino acids, prenlyated (e.g., farnesylated, geranylgeranylated) amino acids, acetylated amino acids, acylated amino acids, PEGylated amino acids, biotinylated amino acids, carboxylated amino acids, phosphorylated amino acids, etc. References to guide the artisan in amino acid modifications are found throughout the various literatures. The experimental protocol can be found in Walker (1998) Protein Protocols On Cd-Rom, Humana Press, Towata, NJ. The modified amino acid may be selected from glycosylated amino acids, PEGylated amino acids, farnesylated amino acids, acetylated amino acids, biotinylated amino acids, amino acids linked to lipid groups, and amino acids linked to organic derivatizing agents.

此外，可通过与多聚体共价连接的方式对抗体进行化学修饰，以例如增加其循环半衰期。作为示例的多聚体和将其连接到肽上的方法在例如US 4,766,106、US 4、179,337、US 4,495,285和US 4,609,546中有所描述。其它示例的多聚体包括聚氧乙烯化多羟基化合物和聚乙二醇(PEG)(例如，分子量在约1,000和约40,000之间，譬如在约2000和约20,000间，例如，约3,000-12,000的PEG)。In addition, antibodies can be chemically modified by covalent attachment to multimers, eg, to increase their circulating half-life. Exemplary polymers and methods of linking them to peptides are described, for example, in US 4,766,106, US 4,179,337, US 4,495,285 and US 4,609,546. Other exemplary polymers include polyoxyethylated polyols and polyethylene glycol (PEG) (e.g., PEG having a molecular weight between about 1,000 and about 40,000, such as between about 2000 and about 20,000, e.g., about 3,000-12,000 ).

在一个技术方案中，本发明提供与第二个选自放射性核、酶、酶底物、辅因子、荧光标记、化学发光标记、肽标签或磁性颗粒的分子连接的CD38BP。在一个技术方案中，CD38BP可与一个或多个抗体片段、核酸(寡核苷酸)、核酸酶、激素、免疫调节物、螯合剂、硼化合物、光敏试剂、染料等连接。这些和其它合适的试剂可直接或间接与本发明的CD38BPs偶联。与第二种试剂间接偶联的例子是通过间隔基团偶联。这些间隔可以是不溶的或可溶的(参见，例如Diener et al.，Science 231.148(1986))，并可选择间隔从而使药物在靶位点和/或在特定条件下从CD38BP释放。可与CD38BPs偶联的治疗试剂的其它实施例包括凝集素和荧光肽。In one embodiment, the present invention provides CD38BP linked to a second molecule selected from radionuclei, enzymes, enzyme substrates, cofactors, fluorescent labels, chemiluminescent labels, peptide tags or magnetic particles. In a technical solution, CD38BP can be linked with one or more antibody fragments, nucleic acid (oligonucleotide), nuclease, hormone, immune modulator, chelating agent, boron compound, photosensitizer, dye, etc. These and other suitable reagents can be conjugated directly or indirectly to the CD38BPs of the invention. An example of indirect coupling to a second reagent is through a spacer group. These spacers can be insoluble or soluble (see, eg, Diener et al., Science 231.148 (1986)), and can be chosen so that the drug is released from the CD38BP at the target site and/or under specific conditions. Other examples of therapeutic agents that can be conjugated to CD38BPs include lectins and fluorescent peptides.

在一个技术方案中，提供了含一个或多个放射性标记的氨基酸的CD38BP衍生物。放射性标记的CD38BP可用于诊断和治疗目的(与放射性分子的偶联是另一个可能的特征)。标记多肽的非限制性例子包括 但不限定于³H、¹⁴C、¹⁵N、³⁵S、⁹⁰Y、⁹⁹Tc、¹²⁵I、¹³¹I和¹⁸⁶Re。制备放射性标记的氨基酸和相关的肽衍生物的方法是本领域已知的(参见，例如Junghans etal.，in Cancer Chemotherapy and Biotherapy 655-686(2dedition，Chafner andLongo，eds.，Lippincott Raven(1996))及US 4,681,581、US 4,735,210、US 5,101,827、US5,102,990(US RE35,500)、US5,648,471和US 5,697,902。例如，放射性同位素可通过氯胺T方法偶联。In one embodiment, there is provided a CD38BP derivative comprising one or more radiolabeled amino acids. Radiolabeled CD38BP can be used for diagnostic and therapeutic purposes (conjugation to radioactive molecules is another possible feature). Non-limiting examples of labeled polypeptides include, but are not limited to, ³ H, ¹⁴ C, ¹⁵ N, ³⁵ S, ⁹⁰ Y, ⁹⁹ Tc, ¹²⁵ I, ¹³¹ I, and ¹⁸⁶ Re. Methods for preparing radiolabeled amino acid and related peptide derivatives are known in the art (see, e.g., Junghans et al., in Cancer Chemotherapy and Biotherapy 655-686 (2dedition, Chafner and Longo, eds., Lippincott Raven (1996)) and US 4,681,581, US 4,735,210, US 5,101,827, US 5,102,990 (US RE35,500), US 5,648,471 and US 5,697,902. For example, radioisotopes can be coupled by the chloramine T method.

在诊断中有用的放射性核是铟放射性同位素，用于治疗的是细胞毒的钇放射性同位素。通常发射质光子的放射性同位素在诊断(放射免疫显像(RIS))方法上是有用的。俄歇电子具有很短的径长(5-10nm)，需要使其内在化而产生细胞毒性(参见，例如Adelstein etal.，Nucl.Med.Biol.14，165-169(1987))。因此，偶联这种放射性同位素的肽可用于诊断方法，但仅有内在化的肽被认为可释放治疗意义上的俄歇电子。α颗粒需要与细胞紧邻(在3-4个细胞直径之内)，才能作为有效的治疗试剂(Vriesendorp et al.，″Radioimmunoglobulin therapy，″in High DoseCancer Therapy Armitage et al.，(eds).(Williams & Wilkins，Baltimore，Md.1992))。俄歇电子和α反射器由于其短范围发射，所以均可认为具有高度选择性，典型地不照射邻近正常细胞。The radionuclide useful in diagnosis is the indium radioisotope and the cytotoxic yttrium radioisotope in therapy. Radioisotopes that generally emit protons are useful in diagnostic (radioimmunoimaging (RIS)) methods. Auger electrons have a very short path length (5-10 nm) and need to be internalized for cytotoxicity (see, eg, Adelstein et al., Nucl. Med. Biol. 14, 165-169 (1987)). Thus, peptides conjugated to this radioisotope are useful in diagnostic methods, but only internalized peptides are thought to release Auger electrons of therapeutic interest. Alpha particles need to be in close proximity (within 3-4 cell diameters) to cells to be effective therapeutic agents (Vriesendorp et al., "Radioimmunoglobulin therapy," in High Dose Cancer Therapy Armitage et al., (eds).(Williams & Wilkins, Baltimore, Md. 1992)). Auger electrons and alpha reflectors can both be considered highly selective due to their short-range emission, typically not illuminating adjacent normal cells.

射电金属¹¹¹In和⁹⁰Y分别是纯的γ-发射者和纯的β-发射者。最常用的俄歇电子发射者碘-125的半衰期约为60天，通常通过免疫偶联物在体内释放(由于去卤化)。最常用于临床的α发射者砹-211和铋-212具有相对较短的半衰期(分别为7.2h和1.0h)，而且在第一次α发射后衰减为不被免疫偶联物保留的放射性同位素(Wilbur，Antibiot.Immunoconjug.Radiopharm.4，5-97(1991))。对于诊断应用而言，可使用标记铟-111或锝-99m的CD38BPs。这两种放射性同位素均在合适的能量范围(100-250keV)内释放γ射线用于成像。典型地，低于该范围的能量不足以穿透而到达外部成像设备。更高的能量水平难以校准，因此所提供的诊断图像分辨率较差。⁹⁹Tc的短半衰期典型地可限制其在具有快速肿瘤吸收的免疫偶联物中的使用。Radiometal ¹¹¹ In and ⁹⁰ Y are pure γ-emitters and pure β-emitters, respectively. The most commonly used Auger electron emitter, iodine-125, has a half-life of approximately 60 days and is usually released in vivo by immunoconjugates (due to dehalogenation). The most commonly used alpha emitters astatine-211 and bismuth-212 have relatively short half-lives (7.2h and 1.0h, respectively) and decay to radioactivity not retained by the immunoconjugate after the first alpha emission Isotopes (Wilbur, Antibiot. Immunoconjug. Radiopharm. 4, 5-97 (1991)). For diagnostic applications, CD38BPs labeled with indium-111 or technetium-99m can be used. Both radioisotopes emit gamma rays in the appropriate energy range (100-250 keV) for imaging. Typically, energy below this range is insufficient to penetrate to external imaging equipment. Higher energy levels are difficult to calibrate and therefore provide poorer resolution diagnostic images. The short half-life ^of99Tc can typically limit its use in immunoconjugates with rapid tumor uptake.

在一个技术方案中，提供了偶联第一和第二放射性同位素的第一和第二CD38BPs。在另一个技术方案中，提供了具有两个放射性同位素的单个CD38BP。用两个单独的放射性同位素，例如一个用于成像，另一个用于治疗的优势是利于门诊患者的治疗。诊断所用的低量放射性不具 有辐射危险，而治疗性放射性同位素，例如纯β发射者所发射的辐射典型地将大量在靶细胞邻近被吸收。In one embodiment, first and second CD38BPs coupled to first and second radioisotopes are provided. In another embodiment, a single CD38BP with two radioisotopes is provided. The advantage of using two separate radioisotopes, eg one for imaging and the other for therapy, facilitates outpatient treatment. The low levels of radioactivity used in diagnostics pose no radiation hazard, whereas radiation emitted by therapeutic radioisotopes, such as pure beta emitters, will typically be absorbed in large quantities in the vicinity of target cells.

放射性同位素可直接或间接与CD38BP连接。例如，放射性同位素 ¹²⁵I、¹³¹I1、⁹⁹Tc、¹⁸⁶Re和¹⁸⁸Re可通过氨基酸功能基团与蛋白(包括抗体)共价连接。对放射性碘而言，它通常通过酪氨酸的苯基连接。有多种方法来完成这种连接：氯胺-T(参见，例如Greenwood etal.，BiochemJ.89，114-123(1963)及lodogen(Salacinski et al.，Anal.Biochem.HZ，136-146(1981))。Tc和Re放射性同位素可通过半胱氨酸巯基基团共价连接(参见，例如Griffiths et al.，Cancer Res.51，4594-4602(1991))。但这些组分相对更适于诊断目的，因为机体通常会破坏这些共价键，从而向循环系统释放放射性同位素。Radioactive isotopes can be directly or indirectly linked to CD38BP. For example, radioactive isotopes ¹²⁵ I, ¹³¹ I1, ⁹⁹ Tc, ¹⁸⁶ Re and ¹⁸⁸ Re can be covalently linked to proteins (including antibodies) through amino acid functional groups. For radioactive iodine, it is usually attached through the phenyl group of tyrosine. There are several ways to accomplish this linkage: chloramine-T (see, for example Greenwood et al., Biochem J.89, 114-123 (1963) and lodogen (Salacinski et al., Anal. Biochem. HZ, 136-146 ( 1981)). Tc and Re radioisotopes can be covalently linked via cysteine sulfhydryl groups (see, for example, Griffiths et al., Cancer Res.51, 4594-4602 (1991)). But these components are relatively more suitable For diagnostic purposes, as the body normally breaks these covalent bonds, releasing the radioisotope into the circulatory system.

CD38BP也可被用于检测的酶，例如辣根过氧化物酶、β-半乳糖苷酶、荧光素酶、碱性磷酸酶、葡萄糖氧化酶等标记。CD38BP也可用生物素标记，从而可通过亲和素或链亲和素的结合而进行间接检测。CD38BP也可用预测的被第二种报告分子(例如，亮氨酸拉链配对序列、第二种抗体的结合位点、金属结合结构域、抗原决定簇标签等)识别的多肽抗原决定簇标记。其它的酶偶联物候选者的例子包括苹果酸脱氢酶、葡萄球菌核酸酶、delta-V-类固醇异构酶、酵母乙醇脱氢酶、α-磷酸甘油脱氢酶、磷酸丙糖异构酶、天冬氨酰酶、葡萄糖氧化酶、核糖核酸酶、尿素酶、过氧化氢酶、葡萄糖-6-磷酸脱氢酶、葡糖淀粉酶和乙酰胆碱酯酶。CD38BP can also be labeled with enzymes used for detection, such as horseradish peroxidase, β-galactosidase, luciferase, alkaline phosphatase, glucose oxidase, etc. CD38BP can also be labeled with biotin, allowing indirect detection by binding of avidin or streptavidin. CD38BP can also be labeled with a polypeptide epitope predicted to be recognized by a second reporter molecule (eg, leucine zipper pairing sequence, binding site for a second antibody, metal binding domain, epitope tag, etc.). Examples of other enzyme conjugate candidates include malate dehydrogenase, staphylococcal nuclease, delta-V-steroid isomerase, yeast alcohol dehydrogenase, alpha-glycerophosphate dehydrogenase, triose phosphate isomerase enzyme, aspartylase, glucose oxidase, ribonuclease, urease, catalase, glucose-6-phosphate dehydrogenase, glucoamylase, and acetylcholinesterase.

其它的示例的标记基团通常包括但不限定于自旋标记分子和其它具有诊断价值的标记基团。Other exemplary labeling groups generally include, but are not limited to, spin-labeled molecules and other labeling groups of diagnostic value.

在一个技术方案中，本发明提供交联的CD38BP衍生物。例如，这种CD38BP衍生物可提供两个或多个(相同类型或不同类型的，例如，可产生双特异性抗体)、其中至少一个特异/选择性针对CD38的抗体交联而产生。合适的交联物包括那些通过合适的间隔区(例如，m-马来酰亚胺苯甲酸-N-羟基琥珀酰亚胺酯)而具有两个不同的反应基团的异源双功能或同源双功能(例如，二琥珀酰亚胺基底物)物质。这种连接物可从Pierce ChemicalCompany，Rockford，III获得。In one technical solution, the present invention provides cross-linked CD38BP derivatives. For example, such CD38BP derivatives can be produced by cross-linking two or more (of the same type or different types, for example, bispecific antibodies can be produced), at least one of which is specific/selective for CD38. Suitable crosslinkers include those heterobifunctional or homobifunctional with two different reactive groups via a suitable spacer (e.g., m-maleimidebenzoic acid-N-hydroxysuccinimide ester). Source Bifunctional (eg, disuccinimide-based) substances. Such linkers are available from Pierce Chemical Company, Rockford, III.

CD38BPs也可与任何合适的化学基团，例如聚乙二醇(PEG)、甲基或乙烷基团或碳水化合物基团偶联。这些和其它合适的偶联基团可用 于改善CD38BP的生物学特性，例如，增加血清半衰期、溶解性和/或组织结合能力。CD38BPs can also be coupled to any suitable chemical group, such as polyethylene glycol (PEG), methyl or ethylene groups, or carbohydrate groups. These and other suitable coupling groups can be used to improve the biological properties of CD38BP, for example, to increase serum half-life, solubility and/or tissue binding capacity.

CD38BP衍生物可将放射性同位素、蛋白或其它试剂/基团/化合物化学偶联到(a)CD38BP的N末端侧链或C末端侧链或其亚基(例如，抗CD38抗体H链、L链、或抗其CD38特异性/选择性片段)的合适取代基或侧链，或(b)CD38BP的糖链(参见，例如AntibodyEngineeringHandbook，edited by Osamu Kanemitsu，published by Chijin Shokan(1994))。在合适情况下，也可通过在内部残基或糖偶联来产生衍生物。CD38BP derivatives can chemically couple radioisotopes, proteins or other reagents/groups/compounds to (a) N-terminal side chains or C-terminal side chains of CD38BP or subunits thereof (for example, anti-CD38 antibody H chain, L chain , or against a CD38-specific/selective fragment thereof), or (b) a sugar chain of CD38BP (see, for example, Antibody Engineering Handbook, edited by Osamu Kanemitsu, published by Chijin Shokan (1994)). Derivatives may also be produced by coupling internal residues or sugars where appropriate.

CD38BPs也可用检测试剂，例如包括荧光素、荧光素硫氰酸盐、若丹明、5-二甲胺-1-萘磺酰氯化物、镧系元素磷等在内的荧光化合物来衍生。合适的荧光标记的其它实施例包括¹²⁵Eu标记、异硫氰酸盐标记、藻红蛋白标记、藻蓝蛋白标记、异藻蓝蛋白标记、邻苯二甲醛标记、荧光胺标记等。化学发光标记的实施例包括鲁米诺标记、异鲁米诺标记、芳香丫啶酯标记、咪唑标记、丫啶盐标记、草酸酯标记、荧光素标记、荧光素酶标记、发光蛋白质标记等。CD38BPs can also be derivatized with detection reagents such as fluorescent compounds including fluorescein, fluorescein thiocyanate, rhodamine, 5-dimethylamine-1-naphthalenesulfonyl chloride, lanthanide phosphorus, and the like. Other examples of suitable fluorescent labels include ¹²⁵ Eu labels, isothiocyanate labels, phycoerythrin labels, phycocyanin labels, isophycocyanin labels, o-phthalaldehyde labels, fluorescamine labels, and the like. Examples of chemiluminescent labels include luminol labeling, isoluminol labeling, aromatic acridine labeling, imidazole labeling, acridinium salt labeling, oxalate labeling, fluorescein labeling, luciferase labeling, luminescent protein labeling, etc. .

在一个技术方案中，CD38BP衍生物含有偶联的核酸或核酸相关的分子。在本发明的一个方面，偶联的核酸是细胞毒核糖核酸酶。在一个技术方案中，偶联的核酸是反义核酸(例如诊断反义分子的S100A10，它也可是本发明的组合组分或联合给药方法中的单独成分-参见，例如Zhang et al.，J Biol Chem.279(3)，2053-62(2004))。在一个技术方案中，偶联的核酸是抑制性RNA分子(例如siRNA分子)。在一个技术方案中，偶联的核酸是免疫激活核酸(例如，免疫激活的含CpG元件的DNA分子)。在一个技术方案中，偶联的核酸是编码肿瘤抑制基因、抗癌疫苗、抗癌细胞因子或凋亡试剂表达的表达盒。这种衍生物也可含有编码一种或多种诸如植物和细菌毒素这样的细胞毒性蛋白表达的核酸偶联物。In one embodiment, the CD38BP derivative contains a conjugated nucleic acid or nucleic acid-related molecule. In one aspect of the invention, the conjugated nucleic acid is a cytotoxic ribonuclease. In one embodiment, the conjugated nucleic acid is an antisense nucleic acid (e.g. S100A10 of a diagnostic antisense molecule, which may also be a single component of a combination or combination method of the invention - see, e.g., Zhang et al., J Biol Chem. 279(3), 2053-62(2004)). In one embodiment, the conjugated nucleic acid is an inhibitory RNA molecule (eg, an siRNA molecule). In one embodiment, the conjugated nucleic acid is an immune-activating nucleic acid (eg, an immune-activating CpG element-containing DNA molecule). In one technical solution, the conjugated nucleic acid is an expression cassette encoding the expression of a tumor suppressor gene, an anticancer vaccine, an anticancer factor or an apoptotic agent. Such derivatives may also contain nucleic acid conjugates encoding the expression of one or more cytotoxic proteins such as plant and bacterial toxins.

在一个技术方案中，CD38BP与功能性核酸分子偶联。功能性核酸分子包括反义分子、干扰核酸分子(例如，siRNA分子)、核酸适体、核酶、三螺旋分子及外源性指导序列。功能性核酸分子可作为靶分子特异活性的效应物、抑制物、调节物及刺激物，或功能性核酸分子可具有独立于任何其它分子的从头合成活性。用于设计及使用反义核酸分子的 方法和技术的代表性例子可在以下非限制性美国专利列表中找到：US5,135,917、US 5,294,533、US5,627,158、US 5,641,754、US 5,691,317、US 5,780,607、US 5,786,138、US 5,849,903、US5,856,103、US 5,919,772、US 5,955,590、US 5,990,088、US 5,994,320、US 5,998,602、US6,005,095、US 6,007,995、US 6,013,522、US 6,017,898、US 6,018,042、US 6,025,198、US6,033,910、US 6,040,296、US 6,046,004、US 6,046,319和US6,057,437。In one technical solution, CD38BP is coupled to a functional nucleic acid molecule. Functional nucleic acid molecules include antisense molecules, interfering nucleic acid molecules (eg, siRNA molecules), aptamers, ribozymes, triple helix molecules, and exogenous guide sequences. Functional nucleic acid molecules can act as effectors, inhibitors, modulators, and stimulators of the specific activity of a target molecule, or functional nucleic acid molecules can have de novo synthesis activity independent of any other molecule. Representative examples of methods and techniques for designing and using antisense nucleic acid molecules can be found in the following non-limiting list of U.S. patents: US 5,135,917; US 5,294,533; US 5,627,158; US 5,641,754; 5,786,138、US 5,849,903、US5,856,103、US 5,919,772、US 5,955,590、US 5,990,088、US 5,994,320、US 5,998,602、US6,005,095、US 6,007,995、US 6,013,522、US 6,017,898、US 6,018,042、US 6,025,198、US6,033,910、US 6,040,296、 US 6,046,004, US 6,046,319 and US 6,057,437.

在一个技术方案中，CD38BP与核酸适体偶联。核酸适体是，例如以特定方式，与靶分子相互作用的分子。典型地，核酸适体是长度为15-50个碱基的小核酸，可折叠成诸如茎-环或G-四聚体这样的确定的二级和三级结构。核酸适体可与诸如ATP(US 5,631,146)和茶碱(US5,580,737)这样的小分子及诸如逆转录酶(US 5,786,462)和凝血酶(US5,543,293)这样的大分子结合。如何生成和使用与多种不同的靶分子结合的核酸适体的代表性实施例可在以下非限制性美国专利列表中找到：US 5,476,766、US 5,503,978、US 5,631,146、US5,731,424、US5,780,228、US 5,792,613、US 5,795,721、US 5,846,713、US 5,858,660、US5,861,254、US 5,864,026、US 5,869,641、US 5,958,691、US 6,001,988、US 6,011,020、US6,013,443、US 6,020,130、US 6,028,186、US 6,030,776和US 6,051,698。In one technical solution, CD38BP is coupled to a nucleic acid aptamer. Aptamers are molecules that, for example, interact with a target molecule in a specific manner. Typically, aptamers are small nucleic acids 15-50 bases in length that fold into defined secondary and tertiary structures such as stem-loop or G-tetramer. Aptamers can bind small molecules such as ATP (US 5,631,146) and theophylline (US 5,580,737) and macromolecules such as reverse transcriptase (US 5,786,462) and thrombin (US 5,543,293). Representative examples of how to generate and use nucleic acid aptamers that bind to a variety of different target molecules can be found in the following non-limiting list of U.S. patents: US 5,476,766; US 5,503,978; US 5,631,146; US 5,731,424; US 5,792,613、US 5,795,721、US 5,846,713、US 5,858,660、US5,861,254、US 5,864,026、US 5,869,641、US 5,958,691、US 6,001,988、US 6,011,020、US6,013,443、US 6,020,130、US 6,028,186、US 6,030,776和US 6,051,698。

在一个技术方案中，本发明提供与核酶偶联的CD38BP。核酶是能催化分子内或分子间化学反应的核酸分子。因此，核酶是具有催化功能的核酸。有多种不同类型的，催化核酸酶或核酸聚合酶类型反应的核酶，这些反应基于天然系统中发现的核酶，譬如(a)锤头型核酶(例如，描述在US 5,334,711、US 5,436,330、US 5,616,466、US 5,633,133、US 5,646,020、US 5,652,094、US 5,712,384、US 5,770,715、US 5,856,463、US 5,861,288、US 5,891,683、US 5,891,684、US 5,985,621、US5,989,908、US 5,998,193、US 5,998,203、WO9858058、WO 9858057和WO 9718312中)。(b)发卡型核酶(例如，描述在US 5,631,115、US 5,646,031、US 5,683,902、US 5,712,384、US 5,856,188、US 5,866,701、US 5,869,339和US6,022,962中)，及(c)四膜虫型核酶(例如，描述在US 5,595,873和US 5,652,107中)。还有很多不是在天然系统中发现的核酶，但它们可被构建，以从头催化特定反应(这类实施例在例如 US 5,580,967、US 5,688,670、US 5,807,718和US 5,910,408中描述)。典型地，核酶剪切RNA或DNA底物，更通常是剪切RNA底物。典型地，核酶通过识别并结合到靶底物上及随后的切割来剪切核酸底物。该识别过程主要是基于典型的或非典型的碱基配对相互作用。因为靶底物的识别基于靶底物的序列，所以该特性使得核酶成为核酸靶特异性剪切的非常优秀的候选者。如何生成并使用核酶以催化多种不同的反应的代表性实施例可在以下非限制性美国专利列表中找到：US 5,646,042、US5,693,535、US 5,731,295、US 5,811,300、US5,837,855、US 5,869,253、US 5,877,021、US 5,877,022、US 5,972,699、US 5,972,704、US5,989,906和US 6,017,756。In one technical solution, the present invention provides CD38BP coupled with ribozyme. Ribozymes are nucleic acid molecules that catalyze intramolecular or intermolecular chemical reactions. Thus, ribozymes are nucleic acids with catalytic function. There are several different types of ribozymes that catalyze nuclease or nucleic acid polymerase type reactions based on ribozymes found in natural systems such as (a) hammerhead ribozymes (described for example in US 5,334,711, US 5,436,330 、US 5,616,466、US 5,633,133、US 5,646,020、US 5,652,094、US 5,712,384、US 5,770,715、US 5,856,463、US 5,861,288、US 5,891,683、US 5,891,684、US 5,985,621、US5,989,908、US 5,998,193、US 5,998,203、WO9858058、WO 9858057和WO 9718312). (b) hairpin-type ribozymes (for example, described in US 5,631,115, US 5,646,031, US 5,683,902, US 5,712,384, US 5,856,188, US 5,866,701, US 5,869,339 and US 6,022,962), and (c) Tetrahymena-type ribozymes ( For example, described in US 5,595,873 and US 5,652,107). There are also many ribozymes that are not found in natural systems, but which can be engineered to catalyze specific reactions de novo (such examples are described, for example, in US 5,580,967, US 5,688,670, US 5,807,718, and US 5,910,408). Typically, ribozymes cleave RNA or DNA substrates, more usually RNA substrates. Typically, ribozymes cleave a nucleic acid substrate by recognizing and binding to a target substrate and subsequent cleavage. The recognition process is mainly based on typical or atypical base-pairing interactions. This property makes ribozymes excellent candidates for target-specific cleavage of nucleic acids because the recognition of the target substrate is based on the sequence of the target substrate. Representative examples of how to make and use ribozymes to catalyze a variety of different reactions can be found in the following non-limiting list of U.S. patents: US 5,646,042; US 5,693,535; US 5,731,295; US 5,811,300; US 5,877,021, US 5,877,022, US 5,972,699, US 5,972,704, US 5,989,906 and US 6,017,756.

在一个技术方案中，本发明提供与三螺旋形式功能性核酸偶联的CD38BP。这种核酸分子可与双链或单链核酸相互作用。当三螺旋分子与靶区域相互作用时，形成了被称为三螺旋的结构，其中3条DNA链形成了基于Watson-Crick和Hoogsteen碱基配对的复合物。三螺旋分子可以高亲和性和特异性与靶区域结合。如何生成并使用与多种不同的靶分子相结合的三螺旋形式分子的代表性实施例可在以下非限制性美国专利列表中找到：US 5,176,996、US 5,645,985、US 5,650,316、US5,683,874、US 5,693,773、US 5,834,185、US 5,869,246、US 5,874,566和US 5,962,426。In one technical solution, the present invention provides CD38BP coupled to functional nucleic acid in triple helix form. Such nucleic acid molecules can interact with double-stranded or single-stranded nucleic acids. When a triple-helical molecule interacts with a target region, a structure known as a triple helix is formed, in which the three DNA strands form a complex based on Watson-Crick and Hoogsteen base pairing. Triple helix molecules can bind to target regions with high affinity and specificity. Representative examples of how to make and use molecules in triple helix form that bind to a variety of different target molecules can be found in the following non-limiting list of US patents: US 5,176,996, US 5,645,985, US 5,650,316, US 5,683,874, US 5,693,773 , US 5,834,185, US 5,869,246, US 5,874,566 and US 5,962,426.

在一个技术方案中，CD38BP与外源性指导序列偶联。外源性指导序列(EGSs)是与形成可被切割靶分子的Rnase P所识别的复合物的靶核酸分子相结合的分子。EGSs可被设计为特异性针对所选择的RNA分子。RNAse P在细胞中帮助转运RNA(tRNA)的加工。通过使用可使靶RNA:EGS复合物模拟为天然tRNA底物的EGS，可募集细菌RNAse P来实质切割任何RNA序列(参见，例如WO 92/03566和Forster andAltman，Science 238，407-409(1990)的讨论)。如何生成并使用EGS分子以促进多种不同靶分子剪切的代表性实施例可在以下非限制性美国专利列表中提供：US 5,168,053、US 5,624,824、US 5,683,873、US5,728,521、US 5,869,248和US 5,877,162。In one technical solution, CD38BP is coupled to an exogenous guide sequence. Exogenous guide sequences (EGSs) are molecules that bind to target nucleic acid molecules forming a complex that is recognized by RNase P, which cleaves the target molecule. EGSs can be designed to be specific for selected RNA molecules. RNAse P aids in the processing of transfer RNA (tRNA) in cells. Bacterial RNAse P can be recruited to cleave virtually any RNA sequence by using EGS, which allows the target RNA:EGS complex to mimic a natural tRNA substrate (see, e.g., WO 92/03566 and Forster and Altman, Science 238, 407-409 (1990). )discussion). Representative examples of how to generate and use EGS molecules to facilitate cleavage of a variety of different target molecules are provided in the following non-limiting list of U.S. patents: US 5,168,053, US 5,624,824, US 5,683,873, US 5,728,521, US 5,869,248, and US 5,877,162 .

在一个技术方案中，CD38BP可与诸如siRNA或其它RNAi分子(例如，约20-25个核苷酸的抑制性双链(ds)RNA分子)这样的干扰核酸分子偶联，它可针对并干扰诸如参与CD38介导的疾病或症状这样的基 因表达产物的靶基因表达产物的作用。在例如Nishikura，Cell.107(4).415-8(2001)、Fjose et al.，Biotechnol Annu Rev.7，31-57(2001)、Hanon，Nature 418，244-51(2002)、Brantl，Biochim Biophys Acta.1575(1-3)，15-25(2002)、Tuschl，Chembiochem.2(4)，239-45(2001)、Caplen，Expert Opin Biol Ther.3(4)，575-86(2003)、Lu et al.，CurrOpin MoI Ther.5(3)，225-34(2003)、Shuey et al.，Drug DiscovToday.7(20)，1040-6(2002)、Shi，Trends Genet.19(1)，9-12(2003)、Kovar et al.，SeminCancer Biol.13(4)，275-81(2003)、Lavrey et al.，Curr Opin Drug Discov Devel.6(4)，561-9(2003)、Clewey，CommunDis Public Health.6(2)，162-3(2003)、Duxbury etal.，J Surg Res.117(2)，339-44(2004)、Caplen et al.，Ann N Y Acad Sci.1002，56-62(2003)、WO 01/75164、US 6,506,559、US 20040086884、US20040077574、US 20040063654、US 20040033602、US 20030167490、US 20030157030、US 20030114409、US 20030108923、US20040014113和US 20020132788中提供了产生和使用干扰核酸分子的方法。In one technical solution, CD38BP can be conjugated to interfering nucleic acid molecules such as siRNA or other RNAi molecules (e.g., inhibitory double-stranded (ds) RNA molecules of about 20-25 nucleotides), which can target and interfere with Role of target gene expression products such as gene expression products involved in CD38-mediated diseases or conditions. In eg Nishikura, Cell. 107(4).415-8(2001), Fjose et al., Biotechnol Annu Rev.7, 31-57(2001), Hanon, Nature 418, 244-51(2002), Brantl, Biochim Biophys Acta.1575(1-3), 15-25(2002), Tuschl, Chembiochem.2(4), 239-45(2001), Caplen, Expert Opin Biol Ther.3(4), 575-86( 2003), Lu et al., CurrOpin MoI Ther.5(3), 225-34(2003), Shuey et al., Drug DiscovToday.7(20), 1040-6(2002), Shi, Trends Genet.19 (1), 9-12(2003), Kovar et al., Semin Cancer Biol.13(4), 275-81(2003), Lavrey et al., Curr Opin Drug Discov Devel.6(4), 561-9 (2003), Clewey, CommunDis Public Health.6(2), 162-3(2003), Duxbury et al., J Surg Res. 117(2), 339-44(2004), Caplen et al., Ann N Y Acad Sci.1002，56-62(2003)、WO 01/75164、US 6,506,559、US 20040086884、US20040077574、US 20040063654、US 20040033602、US 20030167490、US 20030157030、US 20030114409、US 20030108923、US20040014113和US 20020132788中提供了产生and methods of using interfering nucleic acid molecules.

在一个技术方案中，CD38BP可与肿瘤靶向结构域肽或分子偶联。在一个技术方案中，CD38BP与肿瘤靶向因子VII序列偶联。In one technical solution, CD38BP can be coupled to a tumor targeting domain peptide or molecule. In one technical solution, CD38BP is coupled to the sequence of tumor targeting factor VII.

用于将CD38BP与偶联分子相偶联的本领域中任何已知的方法，譬如如上所述的方法，均可使用，包括在Hunter et al.，Nature.144，945(1962)、David et al.，Biochemistry 13，1014(1974)、Pain et al.，J.Immunol.Meth.40，219(1981)和Nygren，J.Histochem.and Cytochem.30，407(1982)中所述的方法。可以任何合适的方式来实现连接/偶联。例如，共价连接可采用二硫键的形式(如果需要且合适的情况下)，可将CD38BP构建为含有额外的半胱氨酸密码子，但需要它不干扰分子的CD38结合活性。具有修饰的CD38BP中半胱氨酸巯基基团反应性的毒素分子可与CD38BP肽形成免疫连接物。选择性地，可使用固相多肽技术直接向CD38BP中引入巯基基团。例如，Hiskey，Peptides 3，137(1981)描述了向肽中引入巯基的方法。在Maasen et al.，Eur.J.Biochem.134，32(1983)中描述了向蛋白中引入巯基的方法。当存在正确的巯基基团时，可纯化细胞毒素和CD38BP，二者的硫基被还原；细胞毒素和配体相混合(例如，以约1∶5到1∶20的比例)；在室温下可完成二硫键的形成(通常约20到30分钟)。然后，可通过磷酸缓冲盐透析或诸如葡聚 糖这样的树脂进行层析来除去混合物中未反应的配体及毒素分子。Any method known in the art for conjugating CD38BP to a coupling molecule, such as those described above, can be used, including in Hunter et al., Nature. 144, 945 (1962), David et al. al., Biochemistry 13, 1014 (1974), Pain et al., J. Immunol. Meth. 40, 219 (1981) and Nygren, J. Histochem. and Cytochem. 30, 407 (1982). Linking/coupling can be accomplished in any suitable manner. For example, covalent linkage can take the form of a disulfide bond (if desired and appropriate), and CD38BP can be constructed to contain an additional cysteine codon, but only if it does not interfere with the CD38 binding activity of the molecule. Toxin molecules with modified cysteine sulfhydryl group reactivity in CD38BP can form immunoconjugates with CD38BP peptides. Alternatively, sulfhydryl groups can be introduced directly into CD38BP using solid phase peptide technology. For example, Hiskey, Peptides 3, 137 (1981) describes a method for introducing sulfhydryl groups into peptides. A method for introducing sulfhydryl groups into proteins is described in Maasen et al., Eur. J. Biochem. 134, 32 (1983). When the correct sulfhydryl group is present, cytotoxin and CD38BP can be purified with their thio groups reduced; cytotoxin and ligand are mixed (e.g., in a ratio of about 1:5 to 1:20); at room temperature Disulfide bond formation can be completed (usually about 20 to 30 minutes). Unreacted ligand and toxin molecules can then be removed from the mixture by phosphate buffered saline dialysis or chromatography on a resin such as dextran.

可通过细胞毒性组分上的反应基团或通过使用交联试剂将多种类型的细胞毒性组分加入到蛋白中。常用的可在体内与胺形成稳定共价键的反应基团是异硫氰酸盐(Meanset al.，Chemical modifications ofproteins(Holden-Day，San Francisco 1971)pp.105-110)。该基团优选地与赖氨酸的ε-氨基反应。顺丁烯二酰亚胺通常用作反应基团以在体内与半胱氨酸上的巯基基团形成稳定的共价键(Ji.，Methods Enzymol 91.，580-609(1983))。典型地，单克隆抗体不能与射电金属离子形成共价键，但它们间接通过使用与抗体共价连接的螯合剂来吸附在抗体上。螯合剂可通过氨基酸残基的氨基基团(Meares etal.，Anal.Biochem..142，68-78(1984))及巯基基团(Koyama，Chem.Abstr.120，217262t(1994))，也可通过碳水化合物基团(Rodwell et al.，PNAS USA 83，2632-2636(1986)、Quadri et al.，Nucl.Med.Biol.20，559-570(1993))连接。由于这些螯合剂含有两种类型的功能基团，一种结合金属离子，另一种参与抗体的螯合，因此它们通常称为双功能螯合剂(Sundberg et al.，Nature 250，587-588(1974))。Various types of cytotoxic components can be incorporated into proteins through reactive groups on the cytotoxic component or through the use of cross-linking reagents. A commonly used reactive group capable of forming stable covalent bonds with amines in vivo is isothiocyanate (Means et al., Chemical modifications of proteins (Holden-Day, San Francisco 1971) pp. 105-110). This group preferably reacts with the ε-amino group of lysine. Maleimide is commonly used as a reactive group to form a stable covalent bond with a sulfhydryl group on cysteine in vivo (Ji., Methods Enzymol 91., 580-609 (1983)). Typically, monoclonal antibodies are unable to form covalent bonds with radiometal ions, but they are adsorbed to antibodies indirectly through the use of chelating agents covalently attached to the antibodies. Chelating agent can pass amino group (Meares et al., Anal.Biochem..142,68-78 (1984)) and sulfhydryl group (Koyama, Chem.Abstr.120,217262t (1994)) of amino acid residue, also Attachment can be via carbohydrate groups (Rodwell et al., PNAS USA 83, 2632-2636 (1986), Quadri et al., Nucl. Med. Biol. 20, 559-570 (1993)). Since these chelators contain two types of functional groups, one that binds metal ions and the other that participates in the chelation of antibodies, they are often referred to as bifunctional chelators (Sundberg et al., Nature 250, 587-588( 1974)).

具有反应性的功能基团的交联试剂可分为同源双功能或异源双功能的。同源双功能交联试剂的实施例包括与巯基基团(Chen et al.，J BiolChem 266，18237-18243(1991))反应的双顺丁烯二酰亚胺(BMH)，及与氨基基团(Browning et al.，J.Immunol.t43，1859-1867(1989))反应的乙烯基乙二醇(succini midylsucciate)(EGS)。异源双功能交联物的实施例包括马来酰亚胺苯甲酸-N-羟基琥珀酰亚胺酯(MBS)(Myers etal.，J.Immunol.Meth.1^1，129-142(1989))。这些方法比较简单，是常用的。Cross-linking reagents with reactive functional groups can be classified as homobifunctional or heterobifunctional. Examples of homologous bifunctional crosslinking reagents include bismaleimide (BMH) reacted with sulfhydryl groups (Chen et al., J BiolChem 266, 18237-18243 (1991)), and bismaleimide (BMH) reacted with amino groups succini midylsucciate (EGS) reacted with a group (Browning et al., J. Immunol. t43, 1859-1867 (1989)). Examples of heterobifunctional crosslinkers include maleimidobenzoic acid-N-hydroxysuccinimide ester (MBS) (Myers et al., J. Immunol. Meth. 191, 129-142 (1989 )). These methods are relatively simple and commonly used.

治疗性或诊断性试剂也可以，或选择性地通过形成二硫键连接在还原性抗体组分的铰链区。作为选择，这种肽可使用诸如3-(2-吡啶二巯基)丙酸N-羟基琥珀酰亚胺酯(SPDP)这样的异源双功能交联物连接到抗体组分上。Yu et al.，Int.J.Cancer 56，244(1994)。这类偶联方法的通用技术在本领域是已知的。参见例如，Wong，Chemistry OfProteinConjugation And Cross-Linking(CRC Press 1991)、Upeslacis et al.，″Modification of Antibodies by Chemical Methods，″In MonoclonalAntibodies：Principles And Applicatiohs，Birch et al.，(eds.)(Wiley- Liss，Inc.1995)、Price，″Production and Characterization of SyntheticPeptide-Derived Antibodies，″inMonoclonal Antibodies：Production，Engineering And Clinical Application，Ritteret al.，(eds.)(CambridgeUniversity Press 1995)。A therapeutic or diagnostic agent can also, or alternatively, be attached to the hinge region of the reducing antibody component by forming a disulfide bond. Alternatively, such peptides can be attached to the antibody component using a heterobifunctional cross-linker such as N-hydroxysuccinimidyl 3-(2-pyridyldithio)propionate (SPDP). Yu et al., Int. J. Cancer 56, 244 (1994). General techniques for such coupling methods are known in the art. See, for example, Wong, Chemistry Of Protein Conjugation And Cross-Linking (CRC Press 1991), Upeslacis et al., "Modification of Antibodies by Chemical Methods," In Monoclonal Antibodies: Principles And Applicatiohs, Birch et al., (eds.) (Wiley- Liss, Inc. 1995), Price, "Production and Characterization of Synthetic Peptide-Derived Antibodies," in Monoclonal Antibodies: Production, Engineering And Clinical Application, Ritter et al., (eds.) (Cambridge University Press 1995).

在某些技术方案中，标记或其它偶联的取代基团通过各种长度的间隔臂与CD38BP的氨基酸序列连接，以减小可能的立体位阻。In some technical solutions, labels or other coupled substituent groups are connected to the amino acid sequence of CD38BP through spacer arms of various lengths to reduce possible steric hindrance.

未标记的CD38BP(s)可与和CD38BP(s)反应的其它标记的抗体(第二抗体)，例如，针对与抗CD38mAbs连接的人免疫球蛋白恒定区的抗体联合使用。选择性地，可直接标记CD38BP。多种标记可用于直接或间接标记CD38BPs，例如标记放射性核、氟石、酶、酶底物、酶辅因子、酶抑制剂、配体(特别是半抗原)等。Unlabeled CD38BP(s) can be used in combination with other labeled antibodies (secondary antibodies) reactive with CD38BP(s), eg, antibodies directed against human immunoglobulin constant regions linked to anti-CD38 mAbs. Alternatively, CD38BP can be directly labeled. A variety of labels can be used to directly or indirectly label CD38BPs, such as labeling radionuclei, fluorspar, enzymes, enzyme substrates, enzyme cofactors, enzyme inhibitors, ligands (especially haptens), etc.

氨基酸序列插入包括氨基和/或羧基末端融合长度为1个残基到含100或更多残基的多肽，及内部序列插入单个或多个氨基酸残基。末端插入的实施例包括具有N末端甲二磺酰残基的抗体或融合抗原决定簇标签的抗体。其它抗体分子的插入变异体包括抗体的N或C末端融合酶或多肽或PEG，从而增加抗体的血清半衰期。这种抗CD38抗体融合蛋白和类似的含CD38BP序列的融合蛋白是本发明的另一个特征。Amino acid sequence insertions include amino and/or carboxyl terminal fusions of 1 residue in length to polypeptides containing 100 or more residues, and internal sequence insertions of single or multiple amino acid residues. Examples of terminal insertions include antibodies with an N-terminal methionyl residue or antibodies fused to an epitope tag. Insertional variants of other antibody molecules include fusion of the N- or C-terminus of the antibody to an enzyme or polypeptide or PEG, thereby increasing the serum half-life of the antibody. Such anti-CD38 antibody fusion proteins and similar fusion proteins containing CD38BP sequences are another feature of the invention.

在一个技术方案中，本发明提供含与治疗性基团，例如细胞毒素、化疗药物、免疫抑制剂或放射性同位素偶联的诸如人抗CD38抗体这样的本发明的CD38BP的分子。这种偶联物此处称为“免疫连接物”，它含有一个或多个称为“免疫毒素”的细胞毒素。In one embodiment, the invention provides molecules comprising a CD38BP of the invention, such as a human anti-CD38 antibody, conjugated to a therapeutic moiety, such as a cytotoxin, chemotherapeutic drug, immunosuppressant or radioisotope. Such conjugates are referred to herein as "immunoconjugates" and contain one or more cytotoxins referred to as "immunotoxins".

细胞毒素或细胞毒素试剂包括任何对细胞有害(例如杀死)的试剂。对于描述这类本领域周知的药物及其作用机制，参见Goodman etal.，Goodman and Gilman′s ThePharmacological Basis Of Therapeutics，8th Ed.，Macmillan Publishing Co.，1990。其它与制备抗体免疫毒素相关的技术在例如Vitetta，Immunol.Today 14，252(1993)和US5,194,594中提供。A cytotoxin or cytotoxic agent includes any agent that is detrimental to (eg, kills) cells. For a description of such art-known drugs and their mechanisms of action, see Goodman et al., Goodman and Gilman's The Pharmacological Basis Of Therapeutics, 8th Ed., Macmillan Publishing Co., 1990. Other techniques related to the preparation of antibody immunotoxins are provided, for example, in Vitetta, Immunol. Today 14, 252 (1993) and US 5,194,594.

合适的形成本发明的免疫连接物的治疗性试剂包括泰素、细胞松弛素B、短杆菌肽D、溴化乙锭、吐根素、丝裂霉素、足叶乙甙、鬼臼噻吩甙、长春新碱、长春碱、秋水仙碱、阿霉素、正定霉素、二氢炭疽菌素二酮、米托蒽醌、放线菌素D、1-脱氢睾酮、糖皮质激素、普鲁卡因、丁卡因、利多卡因、心得安、嘌呤霉素、抗代谢物(譬如氨甲蝶呤、6- -巯基嘌呤、6-硫鸟嘌呤、胞嘧啶阿拉伯糖苷、氟达拉滨、5-氟尿嘧啶、氮烯咪胺、羟基脲、天门冬酰胺酶、吉西他宾、克拉曲滨)、烷基化试剂(譬如二氯甲基二乙胺、thioepa、苯丁酸氮芥、苯丙氨酸氮芥、亚硝脲氮芥(BSNU)、洛莫司汀(CCNU)、环磷酰胺、甲磺酸丁二醇二酯、二溴磷、链脲霉素、氮烯咪胺(DTIC)、甲基苄肼、丝裂霉素C、施铂锭及其它诸如卡铂这样的铂衍生物)、抗生素(譬如更生霉素(以前称为放线菌素)、博来霉素、正定霉素(以前称为道诺霉素)、阿霉素、去甲氧基柔红霉素、光神霉素、加里刹霉素、丝裂霉素、米托蒽醌、普卡霉素、安曲霉素(AMC))、白喉毒素及相关分子(譬如白喉A链及其活性片段以及杂合分子)、蓖麻毒素(譬如蓖麻蛋白A或去糖基化蓖麻蛋白A链毒素)、霍乱毒素、志贺样毒素(SLT-I、SLT-II、SLT-IIV)、LT毒素、C3毒素、志贺毒素、百日咳毒素、破伤风毒素、大豆Bowman-Birk型蛋白酶抑制剂、假单胞菌外毒素、alorin、肥皂草素、莫迪素、甘丙素、相思豆毒素A链、莫迪素A链、α-帚曲菌素、油桐蛋白、康乃馨蛋白、美洲商陆蛋白(PAPI、PAPII及PAP-S)、苦瓜抑制剂、麻疯树毒蛋白、巴豆毒蛋白、sapaonaria officinalis抑制剂、多花白树素、mitogellin、局限曲菌素、酚霉素、及依诺霉素毒素。可如本文其它地方所述的与本发明的CD38BP联合给药的治疗性试剂也可用作偶联本发明的CD38BP的治疗性基团的候选者。例如，药物基团可以是具有所需生物学活性的蛋白或多肽。这种蛋白可包括，例如有酶活性的毒素或其活性片段，例如相思豆毒素、篦麻毒素A、假单胞菌外毒素或白喉毒素；蛋白，例如肿瘤坏死因子或干扰素-γ；或生物应答调节物，诸如例如淋巴因子、白介素-1(IL-1)、白介素-2(IL-2)、白介素-6(IL-6)、粒细胞巨噬细胞集落刺激因子(GM-CSF)、粒细胞集落刺激因子(G-CSF)或其它从线粒体分离的生长因子和凋亡诱导蛋白。在一个技术方案中，细胞毒素试剂是加里刹霉素、⁹⁰Y、¹²⁵I和¹³¹I。与本发明的CD38BP偶联的治疗性细胞毒素的其它例子包括加里刹霉素和duocarmycins。如上所述，不必将药物基团限制为传统的化学治疗试剂。例如，药物基团可以是具有所需生物活性的蛋白或者多肽。这类蛋白可包括，例如，诸如磷酸酯酶类酶，譬如磷酸酯酶C这样的在细胞表面有活性的试剂。Suitable therapeutic agents for forming immunoconjugates of the invention include taxol, cytochalasin B, gramicidin D, ethidium bromide, ipecacine, mitomycin, etoposide, podophylloside , vincristine, vinblastine, colchicine, doxorubicin, daunomycin, dihydroanthracindione, mitoxantrone, actinomycin D, 1-dehydrotestosterone, glucocorticoids, Lucaine, tetracaine, lidocaine, propranolol, puromycin, antimetabolites (such as methotrexate, 6-mercaptopurine, 6-thioguanine, cytosine arabinoside, fludarabine , 5-fluorouracil, dacarbazine, hydroxyurea, asparaginase, gemcitabine, cladribine), alkylating agents (such as dichloromethyldiethylamine, thioepa, chlorambucil, Phenylalanine mustard, nitrosourea mustard (BSNU), lomustine (CCNU), cyclophosphamide, butanediol mesylate, dibromophos, streptozotocin, dacarbazine (DTIC), procarbazine, mitomycin C, platinum and other platinum derivatives such as carboplatin), antibiotics such as dactinomycin (formerly known as actinomycin), bleomycin , daunomycin (formerly known as daunorubicin), doxorubicin, demethoxydaunorubicin, mithramycin, calicheamicin, mitomycin, mitoxantrone, pulkamycin diphtheria toxin, antromycin (AMC)), diphtheria toxin and related molecules (such as diphtheria A chain and its active fragments and hybrid molecules), ricin (such as ricin A or deglycosylated ricin A chain toxin), cholera toxin, Shiga-like toxin (SLT-I, SLT-II, SLT-IIV), LT toxin, C3 toxin, Shiga toxin, pertussis toxin, tetanus toxin, soybean Bowman-Birk type protease inhibitor, Pseudomonas exotoxin, alorin, saporin, modinol, galanin, abrin A chain, modinus A chain, α-buntoxin, tung tree protein, carnation protein, pokeweed protein ( PAPI, PAPII, and PAP-S), bitter melon inhibitors, jatrophin, crotonin, sapaonaria officinalis inhibitors, gentianin, mitogellin, limiterin, phenomycin, and enoxamycin toxin . Therapeutic agents that can be administered in combination with a CD38BP of the invention as described elsewhere herein may also be used as candidates for therapeutic groups to be conjugated to a CD38BP of the invention. For example, a drug moiety can be a protein or polypeptide with the desired biological activity. Such proteins may include, for example, enzymatically active toxins or active fragments thereof such as abrin, ricin A, pseudomonas exotoxin or diphtheria toxin; proteins such as tumor necrosis factor or interferon-gamma; or Biological response modifiers such as, for example, lymphokines, interleukin-1 (IL-1), interleukin-2 (IL-2), interleukin-6 (IL-6), granulocyte macrophage colony stimulating factor (GM-CSF) , granulocyte colony-stimulating factor (G-CSF) or other growth factors and apoptosis-inducing proteins isolated from mitochondria. In one technical solution, the cytotoxic reagents are calicheamicin, ⁹⁰ Y, ¹²⁵ I and ¹³¹ I. Other examples of therapeutic cytotoxins conjugated to CD38BP of the invention include calicheamicins and duocarmycins. As noted above, drug groups need not be limited to traditional chemotherapeutic agents. For example, a drug moiety can be a protein or polypeptide with the desired biological activity. Such proteins may include, for example, agents such as phosphatase-like enzymes, such as phosphatase C, that are active on the surface of cells.

典型地，毒素的裂解部分可与本发明的抗体或抗体片段的Fab片段连接。其它合适的偶联分子包括核糖核酸酶(Rnase)、DNase I、葡萄 球菌外毒素A、美洲商陆抗病毒蛋白、白喉毒素和假单胞菌外毒素。参见，例如Pastan et al.，Cell 47，641(1986)和Goldenberg，Calif.A CancerJournal for Clinicians 44，43(1994)。其它的适于本发明的毒素是本领域技术人员已知的(参见例如US 6,077,499)。Typically, the cleavage portion of the toxin can be linked to the Fab fragment of an antibody or antibody fragment of the invention. Other suitable conjugate molecules include ribonuclease (RNase), DNase I, staphylococcal exotoxin A, pokeweed antiviral protein, diphtheria toxin, and pseudomonas exotoxin. See, eg, Pastan et al., Cell 47, 641 (1986) and Goldenberg, Calif. A Cancer Journal for Clinicians 44, 43 (1994). Other toxins suitable for the present invention are known to those skilled in the art (see eg US 6,077,499).

诸如抗体这样的CD38BPs的偶联物，和这些细胞毒素基团可通过各种双功能蛋白偶联试剂来制备。这种试剂的例子包括SPDP、IT、诸如己二亚胺酸二甲酯HCl这样的亚氨酯双功能衍生物、诸如二琥珀酰氨基底物这样的活性酯、诸如戊二醛这样的醛、诸如二(p-叠氮苯酰)己烷这样的双叠氮化合物、诸如二(p-重氮苯酰)乙二胺这样的双重氮衍生物、诸如亚苄基2，6-二异氰酸酯这样的二异氰酸酯、诸如1，5-二氟-2，4-二硝基苯这样的双活性氟化合物、以及抗有丝分裂试剂(例如，长春新碱、长春碱、紫杉萜、太平洋紫杉醇和长春瑞滨)。Conjugates of CD38BPs, such as antibodies, and these cytotoxic moieties can be prepared by various bifunctional protein conjugation reagents. Examples of such reagents include SPDP, IT, imidoester bifunctional derivatives such as dimethyl adipimidate HCl, active esters such as disuccinamide substrates, aldehydes such as glutaraldehyde, Bis-azide compounds such as bis(p-azidobenzoyl)hexane, bis-nitrogen derivatives such as bis(p-diazobenzoyl)ethylenediamine, such as benzylidene 2,6-diisocyanate diisocyanates, diactive fluorine compounds such as 1,5-difluoro-2,4-dinitrobenzene, and antimitotic agents (e.g., vincristine, vinblastine, docetaxel, paclitaxel, and vinorel coast).

将这种治疗性基团与诸如抗体这样的CD38BPs偶联的技术是已知的，参见，例如Arnon et al.，″Monoclonal Antibodies For lmmunotargetingOf Drugs In CancerTherapy″，in Monoclonal Antibodies And CancerTherapy，Reisfeld et al.，(eds.)、pp.243-56(Alan R.Liss，Inc.1985)、Hellstrom et al.，″Antibodies For DrugDelivery″，in Controlled DrugDelivery(2nd Ed.)，Robinson et al.，(eds.)、pp.623-53(Marcel Dekker，Inc.1987)、Thorpe，″Antibody Carriers Of Cytotoxic Agents InCancerTherapy：A Review″，in Monoclonal Antibodies′84：Biological AndClinicalApplications，Pinchera et al.，(eds.)、pp.475-506(1985)、″Analysis，Results，And Future Prospective Of The Therapeutic Use Of RadiolabeledAntibodyIn Cancer Therapy″，in Monoclonal Antibodies For CancerDetection And Therapy，Baldwin et al.，(eds.)、pp.303-16(AcademicPress 1985)和Thorpe et al.，″ThePreparation And Cytotoxic Properties OfAntibody-Toxin Conjugates″，Immunol.Rev.62，119-58(1982)。Techniques for conjugating such therapeutic groups to CD38BPs such as antibodies are known, see, e.g., Arnon et al., "Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy", in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. , (eds.), pp.243-56 (Alan R. Liss, Inc.1985), Hellstrom et al., "Antibodies For Drug Delivery", in Controlled Drug Delivery (2nd Ed.), Robinson et al., (eds. ), pp.623-53 (Marcel Dekker, Inc. 1987), Thorpe, "Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review", in Monoclonal Antibodies '84: Biological And Clinical Applications, Pinchera et al., (eds.), pp .475-506 (1985), "Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy", in Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al., (eds.), pp.303-16 ( Academic Press 1985) and Thorpe et al., "The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates", Immunol. Rev. 62, 119-58 (1982).

在一个技术方案中，本发明提供与混合毒素偶联的CD38BP。混合的毒素分子是来源于两个不同(典型地多肽)毒素的分子。通常，肽毒素含有一个或多个负责进行真核细胞结合的结构域，至少一个有酶活性的结构域，及至少一个转位结构域。结合和转位结构域分别是细胞识别和毒素进入所需的。已知具有转位结构域的天然蛋白包括白喉毒素、假单胞菌外毒素A和可能的其它肽毒素。白喉毒素和假单胞菌外毒素A 的转位结构域已被描述(参见，例如Hoch et al.，PNAS USA 82，1692(1985)、Colombatti et al.，J.Biol.Chem.26J.，3030(1986)和Deleerset al.，FEBS Lett.160，82(1983))，这种结构域在其它分子中的存在和定位可通过例如Hwang et al.，Cell 48，129(1987)和Gray et al.，PNASUSA 8J.2645(1984)进行的方法进行测定。对这些技术而言，有用的混合毒素杂交物可通过例如将有酶活性的大肠杆菌志贺样毒素的A亚基(Calderwood et al.，PNAS USA 84，4364(1987))与白喉毒素的转位结构域(氨基酸残基从202到460)与针对特定细胞类型的分子融合而形成，如US 5,906,820所述。三部分的杂和体的定位部分可使分子特异地结合到靶细胞上，白喉毒素转位部分可将志贺样毒素有酶活性的A亚基插入到靶细胞中。志贺样毒素有酶活性的部分与白喉毒素一样作用于细胞的蛋白合成机器，抑制蛋白合成，从而杀死靶细胞。In one technical solution, the present invention provides CD38BP coupled to mixed toxins. A mixed toxin molecule is a molecule derived from two different (typically polypeptide) toxins. Typically, a peptide toxin contains one or more domains responsible for eukaryotic cell binding, at least one enzymatically active domain, and at least one translocation domain. Binding and translocation domains are required for cellular recognition and toxin entry, respectively. Natural proteins known to have translocation domains include diphtheria toxin, Pseudomonas exotoxin A and possibly other peptide toxins. The translocation domains of diphtheria toxin and Pseudomonas exotoxin A have been described (see, for example, Hoch et al., PNAS USA 82, 1692 (1985), Colombatti et al., J. Biol. Chem. 26 J., 3030 (1986) and Deleers et al., FEBS Lett.160, 82 (1983)), the presence and location of this domain in other molecules can be ascertained by e.g. Hwang et al., Cell 48, 129 (1987) and Gray et al. al., PNASUSA 8J.2645 (1984) method carried out measurement. For these techniques, useful mixed toxin hybrids can be obtained by, for example, enzymatically active subunit A of E. coli Shiga-like toxin (Calderwood et al., PNAS USA 84, 4364 (1987)) and diphtheria toxin. The amino acid domain (from amino acid residues 202 to 460) is fused to a cell-type-specific molecule, as described in US 5,906,820. The localization part of the three-part hybrid can specifically bind the molecule to the target cell, and the diphtheria toxin translocation part can insert the enzymatically active A subunit of the Shiga-like toxin into the target cell. The enzymatically active part of Shiga-like toxin, like diphtheria toxin, acts on the protein synthesis machinery of cells, inhibits protein synthesis, and kills target cells.

如本发明所述的免疫连接物也可包含放射性同位素，例如碘-131、钇-90或铟-111，从而产生细胞毒性的放射性药物，用于治疗CD38相关的疾病，例如多发性骨髓瘤。Immunoconjugates according to the invention may also contain radioactive isotopes such as iodine-131, yttrium-90 or indium-111 to generate cytotoxic radiopharmaceuticals for the treatment of CD38-associated diseases such as multiple myeloma.

在一个技术方案中，诸如人抗体这样的本发明的CD38BPs与连接物-螯合剂，例如tiuxetan结合，从而使抗体与放射性同位素偶联。In one embodiment, CD38BPs of the invention, such as human antibodies, are conjugated to a linker-chelating agent, such as tiuxetan, thereby conjugating the antibody to a radioisotope.

其它有用的偶联物替代物包括抗肿瘤类维生素A。紫杉烷偶联物(参见，例如Jaimeet al.，Anticancer Res.21.(2A)，1119-28(2001)、施铂锭偶联物、亚油酸偶联物、加里刹霉素偶联物(参见，例如Damle etal.，Curr Opin Pharmacol.3(4)，386-90(2003)、阿霉素偶联物、格尔德霉素偶联物等也可用于促进癌症的治疗(通常参见Trail et al.，CancerImmunol Immunother.52(5)，328-37(2003))。Other useful conjugate substitutes include antineoplastic retinoids. Taxane conjugates (see, e.g., Jaime et al., Anticancer Res. 21. (2A), 1119-28 (2001), platinum platinum conjugates, linoleic acid conjugates, calicheamicin conjugates Drugs (see, for example, Damle et al., Curr Opin Pharmacol. 3(4), 386-90 (2003), doxorubicin conjugates, geldanamycin conjugates, etc. can also be used to promote the treatment of cancer (usually See Trail et al., Cancer Immunol Immunother. 52(5), 328-37 (2003)).

在一个技术方案中，本发明提供针对本发明的抗CD38抗体的第二和抗独特型抗体。第二抗体是指特异，典型地针对抗CD38抗体的抗体。抗独特型(Id)抗体是识别通常与抗体的抗原结合位点相关的独特决定簇的抗体。可通过免疫具有制备抗Id的mAb的，与抗CD38mAb来源相同物种和基因型的动物来制备。典型地，免疫的动物可识别和对免疫抗体的独特型决定簇产生应答，从而产生针对这些独特型决定簇的抗体)抗Id抗体)。这种抗体描述在例如US 4,699,880中。这种抗体是本发明的另一个特征。In one embodiment, the present invention provides a second and anti-idiotypic antibody directed against the anti-CD38 antibody of the present invention. Secondary antibody refers to an antibody that is specific, typically against an anti-CD38 antibody. Anti-idiotypic (Id) antibodies are antibodies that recognize unique determinants normally associated with the antigen-binding site of an antibody. It can be prepared by immunizing an animal of the same species and genotype as the source of the anti-Id mAb. Typically, the immunized animal recognizes and responds to idiotypic determinants of the immunized antibody, thereby producing antibodies (anti-Id antibodies) directed against these idiotypic determinants. Such antibodies are described eg in US 4,699,880. Such antibodies are another feature of the invention.

抗Id抗体也可作为“免疫原”在另一种动物中诱导免疫应答，从而 产生所谓的抗抗Id抗体。抗抗Id可与原始的诱导抗Id的mAb具有相同的抗原决定簇。因此，通过使用针对mAb的独特型决定簇的抗体可鉴定其它具有相同特异性的克隆表达抗体。抗Id抗体可用任何合适的技术，例如本文其它地方描述的关于本发明的抗CD38抗体和其它CD38BPs的方法进行改变(从而产生抗Id抗体变异体)和/或衍生。例如，抗Id mAbs可与诸如钥孔血蓝蛋白(KLH)这样的载体偶联，并用于免疫BALB/c小鼠。来自这些小鼠的血清典型地含有具有与原始/母CD38抗体即使不同但相似的结合特性的抗抗Id抗体。Anti-Id antibodies can also be used as "immunogens" to induce an immune response in another animal, thereby producing so-called anti-anti-Id antibodies. The anti-anti-Id may have the same epitope as the original anti-Id-inducing mAb. Thus, by using antibodies directed against idiotypic determinants of mAbs, other clones expressing antibodies with the same specificity can be identified. Anti-Id antibodies can be altered (thus generating anti-Id antibody variants) and/or derivatized using any suitable technique, eg, as described elsewhere herein with respect to anti-CD38 antibodies and other CD38BPs of the invention. For example, anti-Id mAbs can be conjugated to a carrier such as keyhole limpet hemocyanin (KLH) and used to immunize BALB/c mice. Sera from these mice typically contain anti-anti-Id antibodies with similar, if different, binding properties to the original/parent CD38 antibodies.

在一个技术方案中，本发明提供编码CD38BP的核酸。CD38BP编码核酸可具有任何合适的特征，并含有任何合适的特征或其组合。因此，例如，CD38BP编码核酸可以是DNA、RNA或其杂和体形式，可包括非天然碱基、修饰的骨架(例如，提高核酸稳定性的硫代磷酸化骨架)或包括二者。核酸可含有促进其在靶宿主细胞中所需表达、复制和/或选择的特征。这种特征的例子包括复制组分的起始点、筛选基因组分、启动子组分、增强子元件组分、多腺苷化序列组分、终止组分等。In one technical solution, the present invention provides nucleic acid encoding CD38BP. A CD38BP-encoding nucleic acid may have any suitable characteristics and contain any suitable characteristics or combinations thereof. Thus, for example, a CD38BP-encoding nucleic acid can be in the form of DNA, RNA, or a hybrid thereof, and can include unnatural bases, a modified backbone (eg, a phosphorothioated backbone that increases nucleic acid stability), or both. Nucleic acids may contain features that facilitate their desired expression, replication and/or selection in target host cells. Examples of such features include origin of replication components, selectable gene components, promoter components, enhancer element components, polyadenylation sequence components, termination components, and the like.

在一个技术方案中，本发明提供含CD38BP编码核酸的载体。载体是指促进CD38BP编码核酸表达、CD38BP肽产生、靶细胞转染/转化、CD38BP编码核酸复制、促进核酸稳定性、促进核酸和/或转化/转染细胞的检测，或其它赋予CD38BP编码核酸有用的生物学功能的递送工具。本发明文中的载体可以是任何合适的载体，包括染色体、非染色体和合成的核酸载体(含合适的表达调控元件的核酸序列)。这种载体的例子包括SV40衍生物、细菌质粒、噬菌体DNA、棒状病毒、酵母质粒、来源于质粒和噬菌体DNA组合的载体和病毒核酸(RNA或DNA)载体。在一个技术方案中，CD38BP编码核酸由裸露的DNA或RNA载体组成，包括例如，线性表达元件(如例如Sykes and Johnston，Nat BiotechT7，355-59(1997)所述)，紧密的核酸载体(如例如US 6,077,835和/或WO 00/70087所述)，质粒载体，例如pBR322、pUC 19/18或pUC118/119，“小”的核酸载体(如例如Schakowski et al.，MoI Ther 3，793-800(2001)所述)，或作为沉淀的核酸载体构建体，譬如CaPO₄沉淀的构建体(如例如WO 00/46147、Benvenisty and Reshef，PNAS USA 83，9551-55(1986)、Wigler et al.，Cell 14，725(1978)、及Coraro and Pearson，Somatic Cell Genetics 7，603(1981)所述)。这类核酸载体及用途在本 领域是周知的(参见例如US 5,589,466和US 5,973,972)。In one technical solution, the present invention provides a vector containing CD38BP-encoding nucleic acid. Carrier refers to promoting the expression of CD38BP-encoding nucleic acid, CD38BP peptide production, target cell transfection/transformation, CD38BP-encoding nucleic acid replication, promoting nucleic acid stability, promoting the detection of nucleic acid and/or transformation/transfection cells, or other useful endowing CD38BP-encoding nucleic acid Biologically functional delivery tool. The vector in the context of the present invention can be any suitable vector, including chromosomal, non-chromosomal and synthetic nucleic acid vectors (nucleic acid sequences containing suitable expression control elements). Examples of such vectors include SV40 derivatives, bacterial plasmids, phage DNA, baculoviruses, yeast plasmids, vectors derived from combinations of plasmids and phage DNA, and viral nucleic acid (RNA or DNA) vectors. In one technical solution, the CD38BP-encoding nucleic acid consists of a naked DNA or RNA vector, including, for example, a linear expression element (as described, for example, in Sykes and Johnston, Nat Biotech T7, 355-59 (1997), a compact nucleic acid vector (such as Such as described in US 6,077,835 and/or WO 00/70087), plasmid vectors, such as pBR322, pUC 19/18 or pUC118/119, "small" nucleic acid vectors (such as for example Schakowski et al., MoI Ther 3, 793-800 (2001) described), or as a precipitated nucleic acid carrier construct, such as a _CaPO Precipitated construct (as for example WO 00/46147, Benvenisty and Reshef, PNAS USA 83, 9551-55 (1986), Wigler et al. , Cell 14, 725 (1978), and Coraro and Pearson, Somatic Cell Genetics 7, 603 (1981)). Such nucleic acid vectors and their use are well known in the art (see eg US 5,589,466 and US 5,973,972).

在一个技术方案中，载体适于在细菌细胞中表达CD38BP。这种载体的实施例包括，例如，指导易于纯化的融合蛋白高水平表达的载体(例如，诸如BlueScript(Stratagene)、pIN载体(Van Heeke & Schuster，JBiol Chem 264，5503-5509(1989)、pET载体(Novagen，Madison Wl)等这样的多功能大肠杆菌的克隆及表达载体)。In one embodiment, the vector is suitable for expressing CD38BP in bacterial cells. Examples of such vectors include, for example, vectors that direct high-level expression of fusion proteins that are readily purified (for example, such as BlueScript (Stratagene), pIN vectors (Van Heeke & Schuster, J Biol Chem 264, 5503-5509 (1989), pET Vectors (Novagen, Madison Wl) and other such multifunctional Escherichia coli cloning and expression vectors).

表达载体也可以或选择性地是适于在酵母系统中进行表达的载体。可使用任何适于在酵母系统中进行表达的载体。例如可用于在酿酒酵母中的合适载体包括，例如，含诸如α因子、乙醇氧化酶及PGH这样的组成型或诱导型启动子的载体(综述可参见：F.Ausubel etal.，ed.Current Protocols in Molecular Biology，Greene Publishing andWileyInterScience New York(1987)、及Grant et al.，Methods in Enzymol 153.516-544(1987))。The expression vector may also or alternatively be a vector suitable for expression in yeast systems. Any vector suitable for expression in yeast systems can be used. For example, suitable vectors for use in S. cerevisiae include, for example, vectors containing constitutive or inducible promoters such as alpha factor, alcohol oxidase, and PGH (for review, see: F. Ausubel et al., ed. Current Protocols in Molecular Biology, Greene Publishing and Wiley InterScience New York (1987), and Grant et al., Methods in Enzymol 153.516-544 (1987)).

核酸和/或载体也可以含有编码分泌/定位序列的核酸序列，这些序列可使诸如新生多肽链这样的多肽定位到所需的细胞内区域、膜或细胞器，或者这些序列可指导多肽分泌到周质空间或细胞培养基中。这类序列在本领域是已知的，包括分泌引导序列或信号肽、细胞器定位序列(例如，核定位序列、ER滞留信号、线粒体穿梭序列、叶绿体穿梭序列)、膜定位/锚定序列(例如，转移终止序列、GPI锚定序列)等。The nucleic acid and/or vector may also contain nucleic acid sequences encoding secretion/localization sequences that allow localization of a polypeptide, such as a nascent polypeptide chain, to desired intracellular regions, membranes, or organelles, or that direct secretion of the polypeptide to peripheral cytoplasmic space or cell culture medium. Such sequences are known in the art and include secretion leader sequences or signal peptides, organelle localization sequences (e.g., nuclear localization sequences, ER retention signals, mitochondrial shuttle sequences, chloroplast shuttle sequences), membrane localization/anchor sequences (e.g. , transfer termination sequence, GPI anchor sequence), etc.

CD3 8BP编码核酸可含有或与任何合适的启动子、增强子及其它促进表达的元件连接。这类元件的实施例包括强表达启动子(例如，人CMV IE启动子/增强子以及RSV、SV40、SL3-3、MMTV和HIV LTR)、有效的多聚(A)终止序列、大肠杆菌中质粒产物的复制起点、用作选择标记的抗生素抗性基因、和/或方便的克隆位点(例如，多连接物)。核酸也可含有与诸如CMV IE这样的组成型启动子相对的诱导型启动子(技术人员指导这类术语是特定条件下基因表达程度的实际描述)。A CD3 8BP-encoding nucleic acid may contain or be linked to any suitable promoter, enhancer, and other expression-promoting elements. Examples of such elements include strong expression promoters (e.g., human CMV IE promoter/enhancer and RSV, SV40, SL3-3, MMTV, and HIV LTRs), efficient poly(A) termination sequences, E. coli An origin of replication for the plasmid product, an antibiotic resistance gene for use as a selectable marker, and/or a convenient cloning site (eg, polylinker). A nucleic acid may also contain an inducible promoter as opposed to a constitutive promoter such as CMV IE (the skilled artisan directs that such terms are an actual description of the extent of gene expression under particular conditions).

在一个技术方案中，核酸可通过病毒载体定位和/或递送到宿主细胞或者宿主动物中。任何合适的病毒载体可用于该目的，有几个是本领域已知的。病毒载体可单独或者与一个或多个病毒蛋白一起，含有任意数目的病毒多核苷酸，这些病毒蛋白可促进本发明的核酸在所需宿主细胞中的递送、复制和/或表达。病毒载体可以是含有全部或部分病毒基因组的多核苷酸、病毒蛋白/核酸偶联物、病毒类似颗粒(V_LP)、与US 5,849, 586和WO 97/04748中所述载体类似的载体、或含有病毒核酸及本发明中核酸的完整的病毒颗粒。病毒颗粒类病毒载体可含有野生型病毒颗粒或修饰的病毒颗粒。病毒载体可以是需要存在另一种载体或野生型病毒来进行复制和/或表达的载体(即，它是辅助依赖病毒)，譬如腺病毒载体扩增子。典型地，这类病毒载体实质上由野生型病毒颗粒，或者其蛋白和/或核酸被修饰以增加转基因能力或促进核酸转染和/或表达的病毒颗粒组成的(这类载体的实施例包括疱疹病毒/AAV扩增子)。典型地，病毒载体类似于、和/或来源于通常感染人类的病毒。在这方面，合适的病毒载体包括，例如，腺病毒载体颗粒(包括任何腺病毒或者来源于腺病毒的病毒)、腺相关病毒载体颗粒(AAV载体颗粒)或其它细小病毒及细小病毒载体颗粒、乳头状瘤病毒载体颗粒、黄质病毒载体、α病毒载体、疱疹病毒载体、痘病毒载体、包括慢病毒载体在内的逆转录病毒载体。这类病毒及病毒载体的实施例参见例如Fields et al.，eds.，VirologyRaven Press，Ltd.，New York(3rd ed.，1996 and 4th ed.，2001)、Encyclopediaof Virology，R.G.Webster et al.，eds.，Academic Press(2nded.，1999)、FundamentalVirology，Fields et al.，eds.，Lippincott-Raven(3rd ed.，1995)、Levine，″Viruses，″Scientific American Library No.37(1992)、Medical Virology，D.O.White et al.，eds.，Acad.Press(2nded.1994)、及Introduction to Modern Virology，Dimock，N.J.etal.，eds.，Blackwell Scientific Publicaions，Ltd.(1994)。In one technical solution, the nucleic acid can be localized and/or delivered into a host cell or host animal by a viral vector. Any suitable viral vector can be used for this purpose, several are known in the art. Viral vectors may contain any number of viral polynucleotides, alone or together with one or more viral proteins that facilitate delivery, replication and/or expression of the nucleic acids of the invention in a desired host cell. Viral vectors can be polynucleotides containing all or part of the viral genome, viral protein/nucleic acid conjugates, virus-like particles (VLP), vectors similar to those described in US _5,849,586 and WO 97/04748, Or complete viral particles containing viral nucleic acids and nucleic acids of the present invention. Virus Particles Virus-like vectors may contain wild-type virus particles or modified virus particles. A viral vector may be one that requires the presence of another vector or a wild-type virus for replication and/or expression (ie, it is a helper-dependent virus), such as an adenoviral vector amplicon. Typically, such viral vectors are substantially composed of wild-type viral particles, or viral particles whose proteins and/or nucleic acids have been modified to increase transgenic capacity or to promote nucleic acid transfection and/or expression (examples of such vectors include Herpesvirus/AAV amplicon). Typically, viral vectors are similar to, and/or derived from, viruses that commonly infect humans. In this regard, suitable viral vectors include, for example, adenoviral vector particles (including any adenovirus or virus derived from adenoviruses), adeno-associated viral vector particles (AAV vector particles) or other parvoviruses and parvoviral vector particles, Papillomavirus vector particles, flavivirus vectors, alphavirus vectors, herpesvirus vectors, poxvirus vectors, retrovirus vectors including lentivirus vectors. Examples of such viruses and viral vectors are found in, for example, Fields et al., eds., VirologyRaven Press, Ltd., New York (3rd ed., 1996 and 4th ed., 2001), Encyclopedia of Virology, RG Webster et al., eds ., Academic Press (2nded., 1999), Fundamental Virology, Fields et al., eds., Lippincott-Raven (3rd ed., 1995), Levine, "Viruses," Scientific American Library No.37 (1992), Medical Virology , DOWhite et al., eds., Acad. Press (2nded. 1994), and Introduction to Modern Virology, Dimock, NJ et al., eds., Blackwell Scientific Publicaions, Ltd. (1994).

可使用本发明中多核苷酸及此处所述方法的病毒载体包括腺病毒和腺相关载体，例如Carter，Curr Opinion Biotech 3，533-539(1992)和Muzcyzka，Curr Top MicrobiolImmunol 158，97-129(1992)。在例如Carter，Contrib.Microbiol.4，85-86(2000)、Smith-Arica，Curr.Cardiol.Rep.3(1)，41-49(2001)、Taj，J.Biomed.Sci.7(4)，279-91(2000)、Vigna et al.，J.Gene Med.2(5)，308-16(2000)、Klimatcheva et al.，Front.Biosci.4，D481-96(1999)、Lever et al.，Biochem.Soc.Trans.27(6)，841-47(1999)、Snyder，J GeneMed.1(3)，166-75(1999)、Gerich et al.，Knee Surg.Sports Traumatol.Arthrosc.5(2)，118-23(1998)、和During，Adv.Drug Deliv.Review 27(1)，83-94(1997)以及US 4,797,368、US 5,139,941、US 5,173,414、US 5,614,404、US5,658,785、US 5,858,775和US 5,994,136中描述了AAV载体的其它类 型与方面。可通过在例如US 4,797,368和Laughlin etal.，Gene 23，65-73(1983)中确定的方法来构建和/或纯化腺相关病毒载体。Viral vectors with which the polynucleotides of the invention and methods described herein can be used include adenoviruses and adeno-associated vectors, such as Carter, Curr Opinion Biotech 3, 533-539 (1992) and Muzcyzka, Curr Top Microbiol Immunol 158, 97-129 (1992). In eg Carter, Contrib.Microbiol.4,85-86(2000), Smith-Arica, Curr.Cardiol.Rep.3(1), 41-49(2001), Taj, J.Biomed.Sci.7(4 ), 279-91(2000), Vigna et al., J. Gene Med.2(5), 308-16(2000), Klimatcheva et al., Front.Biosci.4, D481-96(1999), Lever et al., Biochem. Soc. Trans. 27(6), 841-47(1999), Snyder, J GeneMed. 1(3), 166-75(1999), Gerich et al., Knee Surg. Sports Traumatol. Arthrosc.5(2), 118-23(1998), and During, Adv.Drug Deliv.Review 27(1), 83-94(1997) and US 4,797,368, US 5,139,941, US 5,173,414, US 5,614,404, US5,658,785 Other types and aspects of AAV vectors are described in US 5,858,775 and US 5,994,136. Adeno-associated virus vectors can be constructed and/or purified by the methods established, for example, in US 4,797,368 and Laughlin et al., Gene 23, 65-73 (1983).

另一种类型的使用本发明中多核苷酸和方法的病毒载体是乳头状瘤病毒载体。合适的乳头状瘤病毒载体在本领域是已知的，并在例如中Hewson，MoI Med Today 5(1)，8(1999)、Stephens，Biochem J.248(1)，1-11(1987)和US 5,719,054有所描述。在例如WO99/21979中提供了乳头状瘤病毒载体的实施例。α病毒载体在其它地方可以是基因递送载体α病毒载体在本领域是已知的，并在例如Carter，Curr OpinionBiotech 3，533-539(1992)、Muzcyzka，Curr Top Microbiol Immunol.158，97-129(1992)、Schlesinger，Expert Opin Biol Ther.1(2)，177-91(2001)、Polo et al.，Dev Biol(Basel).104，181-5(2000)、Wahlfors et al.，Gene Ther.7(6)，472-80(2000)、Colombage et al.，Virology.250(1)，151-63(1998)以及WO 01/81609、WO 00/39318、WO 01/81553、WO95/07994和WO 92/10578中有所描述。Another type of viral vector using the polynucleotides and methods of the invention is a papillomavirus vector. Suitable papillomavirus vectors are known in the art and are described, for example, in Hewson, MoI Med Today 5(1), 8 (1999), Stephens, Biochem J. 248(1), 1-11 (1987) and US 5,719,054 as described. Examples of papillomavirus vectors are provided in eg WO99/21979. Alphavirus vectors can be gene delivery vectors elsewhere. Alphavirus vectors are known in the art and described, for example, in Carter, Curr Opinion Biotech 3, 533-539 (1992), Muzcyzka, Curr Top Microbiol Immunol. 158, 97-129 (1992), Schlesinger, Expert Opin Biol Ther.1(2), 177-91(2001), Polo et al., Dev Biol (Basel).104, 181-5(2000), Wahlfors et al., Gene Ther .7(6), 472-80(2000), Colombage et al., Virology.250(1), 151-63(1998) and WO 01/81609, WO 00/39318, WO 01/81553, WO95/07994 and described in WO 92/10578.

另一组病毒载体是疱疹病毒载体。在例如Lachmann et al.，Curr OpinMoI Ther1(5)，622-32(1999)、Fraefel et al.，Adv Virus Res.55，425-51(2000)、Huard et al.，Neuromuscul Z(5)，299-313(1997)、Glorioso et al.，Annu Rev Microbiol.49，675-710(1995)、Latchman，MoI Biotechnol.2(2)，179-95(1994)、和Frenkel et al.，Gene Ther.i(Suppl 1)，S40-6(1994)以及US 6,261,552和US 5,599,691中描述了疱疹病毒载体的实施例。Another group of viral vectors are the herpes virus vectors. In e.g. Lachmann et al., Curr OpinMoI Ther1 (5), 622-32 (1999), Fraefel et al., Adv Virus Res. 55, 425-51 (2000), Huard et al., Neuromuscul Z (5), 299-313 (1997), Glorioso et al., Annu Rev Microbiol. 49, 675-710 (1995), Latchman, MoI Biotechnol. 2 (2), 179-95 (1994), and Frenkel et al., Gene Ther Examples of herpes virus vectors are described in .i (Suppl 1), S40-6 (1994) and US 6,261,552 and US 5,599,691.

包括慢病毒载体在内的逆转录病毒载体在特定环境中也是有利的基因递送工具。在本领域已知有多种逆转录病毒载体。在Miller，Curr TopMicrobiol Immunol 158.1-24(1992)、Salmons and Gunzburg，Human GeneTherapy 4，129-141(1993)、Miller et al.，Methods in Enzvmolosv 217，581-599(1994)、Weber et al.，Curr Opin MoI Ther.3(5)，439-53(2001)、Hu et al.，Pharmacol Rev.52(4)，493-511(2000)、Kim et al.，AdvVirusRes.55，545-63(2000)、PaIu et al.，Rev Med Virol.10(3)，185-202(2000)和Takeuchi et al.，Adv Exp Med Biol.465，23-35(2000)以及US 6,326,195、US 5,888,502、US 5,580,766和US 5,672,510中描述了逆转录病毒载体的实施例。Retroviral vectors, including lentiviral vectors, are also advantageous gene delivery tools in certain circumstances. A variety of retroviral vectors are known in the art. In Miller, Curr Top Microbiol Immunol 158.1-24 (1992), Salmons and Gunzburg, Human GeneTherapy 4, 129-141 (1993), Miller et al., Methods in Enzvmolosv 217, 581-599 (1994), Weber et al., Curr Opin MoI Ther.3(5), 439-53(2001), Hu et al., Pharmacol Rev.52(4), 493-511(2000), Kim et al., AdvVirusRes.55, 545-63( 2000), PaIu et al., Rev Med Virol.10(3), 185-202(2000) and Takeuchi et al., Adv Exp Med Biol.465, 23-35(2000) and US 6,326,195, US 5,888,502, US Examples of retroviral vectors are described in 5,580,766 and US 5,672,510.

腺病毒载体也是基因转移合适的病毒载体。腺病毒载体是本领域已 知的，描述在例如Graham et al，MoI Biotechnol 33(3)，207-220(1995)、Stephenson，Clin DiagnVirol 10(2-3)，187-94(1998)、Jacobs，ClinSci(Lond).85(2)，117-22(1993)、US 5,922,576、US 5,965,358和US 6,168,941和WO98/22588、WO98/56937、WO99/15686、WO99/54441和WO00/32754中。用于本发明的腺病毒载体、疱疹病毒载体和Sindbis病毒载体描述在例如Jolly Cancer Gene Therapy 1，51-64(1994)、Latchman Molec Biotechnol 2，179-195(1994)和Johanninget al.，Nucl Acids Res 23，1495-1501(1995)中。Adenoviral vectors are also suitable viral vectors for gene transfer. Adenoviral vectors are known in the art and described, for example, in Graham et al, MoI Biotechnol 33(3), 207-220 (1995), Stephenson, Clin DiagnVirol 10(2-3), 187-94 (1998), Jacobs , ClinSci (Lond). 85(2), 117-22 (1993), US 5,922,576, US 5,965,358 and US 6,168,941 and WO98/22588, WO98/56937, WO99/15686, WO99/54441 and WO00/32754. Adenovirus vectors, herpesvirus vectors and Sindbis virus vectors used in the present invention are described, for example, in Jolly Cancer Gene Therapy 1, 51-64 (1994), Latchman Molec Biotechnol 2, 179-195 (1994) and Johanning et al., Nucl Acids Res 23, 1495-1501 (1995).

其它合适的病毒载体包括痘病毒载体。这种载体的实施例描述在例如Berencsiet al.，J Infect Dis 183(8)，1171-9(2001)、Rosenwirth etal.，Vaccine 19(13-14)，1661-70(2001)、Kittlesen et al.，J Immunol164(8)，4204-11(2000)、Brown et al.，Gene Ther 7(19)，1680-9(2000)、Kanesa-thasan et al.，Vaccine 19(4-5)、483-91(2000)、Sten，Drua 60(2)，249-71(2000)中。疫苗病毒载体可以是痘病毒载体。这种载体及其用途的实施例在例如Venugopal et al.，Res Vet Sci 57(2)，188-193(1994)、Moss DevBiol Stand 82，55-63(1994)、Weisz et al.，MoI Cell Biol 43，137-159(1994)、Mahrand Payne，Immunobioloev 184(2-3)，126-146(1992)、Hruby，Clin Microbiol Rev3(2)，153-170(1990)和WO92/07944、WO98/13500和WO89/08716中提供。Other suitable viral vectors include poxvirus vectors. Examples of such vectors are described, for example, in Berencsi et al., J Infect Dis 183(8), 1171-9 (2001), Rosenwirth et al., Vaccine 19(13-14), 1661-70 (2001), Kittlesen et al. ., J Immunol164(8), 4204-11(2000), Brown et al., Gene Ther 7(19), 1680-9(2000), Kanesa-thasan et al., Vaccine 19(4-5), 483 -91 (2000), Sten, Drua 60(2), 249-71 (2000). The vaccinia viral vector may be a poxvirus vector. Examples of such vectors and their use are e.g. Biol 43, 137-159(1994), Mahrand Payne, Immunobioloev 184(2-3), 126-146(1992), Hruby, Clin Microbiol Rev3(2), 153-170(1990) and WO92/07944, WO98/ 13500 and WO89/08716.

本发明的其它特征包括例如酵母、细菌和哺乳动物(例如，永生化哺乳动物细胞)这样的含核酸、载体或其组合的重组细胞。例如，在一个技术方案中，本发明提供含稳定整合在细胞基因组中，含表达本发明的CD38BP的编码序列的核酸的细胞。在一个技术方案中，本发明提供含例如质粒、粘粒、噬菌体粒或线性表达元件这样的包含编码表达CD38BP的序列的非整合核酸。Additional features of the invention include recombinant cells, such as yeast, bacteria, and mammals (eg, immortalized mammalian cells), that contain nucleic acids, vectors, or combinations thereof. For example, in one technical solution, the present invention provides a cell containing a nucleic acid expressing the coding sequence of CD38BP of the present invention stably integrated in the genome of the cell. In one technical solution, the present invention provides a non-integrating nucleic acid comprising a sequence encoding and expressing CD38BP, such as a plasmid, cosmid, phagemid or linear expression element.

本发明也提供含任何上述针对本发明的CD38BPs的抗原决定簇部分，例如针对-003和-005及-024的CD38抗原决定簇部分的免疫肽。这种免疫原可在含有活性免疫治疗的方法中用于引发直接的免疫应答。本发明进一步提供含这种CD38免疫原和融合部分序列的融合蛋白，该融合序列可提高融合蛋白半衰期(例如，包括免疫球蛋白结构域序列)；利于融合蛋白检测和/或纯化(通过含例如荧光肽序列、报告酶序列、抗 原决定簇标签、六组氨酸序列等)；促进融合蛋白定位(例如，通过含有特异针对靶细胞上受体的配体或配体一部分)；促进不同免疫应答的诱导(例如，对癌抗原或其免疫原片段应答)是细胞毒试剂；或完成任何其组合(例如，热休克融合蛋白伴侣可增加产生的针对融合蛋白非相似的异源抗原部分的免疫应答，而且也增加融合蛋白的体内半衰期)。融合蛋白也含有一个或多个切割位点，特别是在结构域间。The invention also provides immunizing peptides comprising any of the above epitope portions directed against the CD38BPs of the invention, for example against -003 and -005 and -024. Such immunogens can be used to elicit an immediate immune response in methods involving active immunotherapy. The present invention further provides a fusion protein comprising such a CD38 immunogen and a fusion part sequence that increases the half-life of the fusion protein (for example, including an immunoglobulin domain sequence); facilitates detection and/or purification of the fusion protein (by containing, for example, an immunoglobulin domain sequence); fluorescent peptide sequences, reporter enzyme sequences, epitope tags, hexahistidine sequences, etc.); facilitate localization of fusion proteins (e.g., by including a ligand or part of a ligand specific for a receptor on a target cell); promote differential immune responses Induction (e.g., a response to a cancer antigen or immunogenic fragment thereof) is a cytotoxic agent; or accomplishes any combination thereof (e.g., a heat shock fusion protein partner can increase the immune response generated against a dissimilar heterologous antigenic portion of the fusion protein , but also increase the in vivo half-life of the fusion protein). Fusion proteins also contain one or more cleavage sites, particularly between domains.

这种肽的变异体和这种免疫原肽或免疫原肽变异体的衍生物是本发明的另一个特征(例如，这种CD38免疫原肽衍生物可通过化学偶联、基因融合、非共价相互作用等与其它例如抗体、递送、放射性同位素、细胞毒试剂或控制细胞生长的试剂这样的分子实体连接而修饰)。例如，含CD38抗原决定簇序列的肽模拟表位也可作为疫苗候选者。这种肽也可用于抗CD38抗体的纯化。除了此处所述的B细胞抗原决定簇序列外，这种肽也被构建或选择或选择性地含有一个或多个抗CD38T细胞抗原决定簇。这种抗原决定簇可通过本领域已知的任何合适技术进行鉴定(例如通过使用T细胞抗原决定簇预测软件)。Variants of such peptides and derivatives of such immunogenic peptides or variants of immunogenic peptides are another feature of the invention (for example, such derivatives of CD38 immunogenic peptides may be obtained by chemical coupling, gene fusion, non-cooperative valence interactions etc. in conjunction with other molecular entities such as antibodies, delivery, radioisotopes, cytotoxic agents, or agents that control cell growth). For example, peptide mimotopes containing CD38 epitope sequences may also be vaccine candidates. This peptide can also be used for the purification of anti-CD38 antibodies. Such peptides may also be constructed or selected or optionally contain one or more anti-CD38 T cell epitopes in addition to the B cell epitope sequences described herein. Such epitopes may be identified by any suitable technique known in the art (eg by using T cell epitope prediction software).

在一个技术方案中，本发明提供编码这种免疫原肽的核酸。这种核酸可被合适的载体，例如复制缺陷靶载体(例如，靶核酸载体或复制缺陷的靶腺病毒载体)被递送到宿主中。本发明也提供一种或多种这样的免疫原肽和/或免疫原肽编码核酸的组分。In one technical solution, the present invention provides nucleic acids encoding such immunogenic peptides. Such nucleic acids can be delivered to the host by a suitable vector, such as a replication-deficient targeting vector (eg, a targeting nucleic acid vector or a replication-deficient targeting adenoviral vector). The invention also provides one or more such immunogenic peptides and/or components of immunogenic peptide-encoding nucleic acids.

本发明的CD38BPs包括“内在化”CD38BPs，例如内在化抗体。术语“内在化CD38BP”和“内在化抗体”是指能实质抑制或消除CD38相关肽的生物学活性的CD38BP或抗体。典型地，内在化CD38BP，例如内在化抗CD38抗体可直接或间接抑制CD38的功能，例如酶活性、信号转导、诱导细胞因子表达、诱导增殖或分化或诱导裂解，在程度上比由于给药约等量的-003或-005或-024而使这种细胞的抑制更大。CD38BPs of the invention include "internalizing" CD38BPs, such as internalizing antibodies. The terms "internalizing CD38BP" and "internalizing antibody" refer to CD38BP or an antibody that substantially inhibits or eliminates the biological activity of CD38-related peptides. Typically, internalizing CD38BP, such as internalizing an anti-CD38 antibody, can directly or indirectly inhibit CD38 functions, such as enzymatic activity, signal transduction, induction of cytokine expression, induction of proliferation or differentiation, or induction of lysis, to a greater extent than due to administration of Approximately equal amounts of -003 or -005 or -024 resulted in greater inhibition of this cell.

本发明的CD38BP可对至少部分含在CD38中的一个或多个抗原决定簇具有任何合适的亲和性和/或强度。亲和性是指CD38BP与这种抗原决定簇结合的强度。典型地，亲和性通过解离常数K_d来测定，K_d定义为[Ab]×[Ag]/[Ab-Ag]，其中[Ab-Ag]是抗体-抗原复合物(或CD38BP-抗原复合物)的摩尔浓度，[Ab]是未结合的抗体(或CD38BP)的摩尔浓度，[Ag]是未结合的抗原的摩尔浓度。亲和性常数K_a定义为1/K_d。通过竞争作用测定特异性和亲和性的合适方法可在例如，Harlow et al.， Antibodies：A Laboratory Manual，Cold SpringHarbor Laboratory Press，Cold Spring Harbor，N.Y.，1988)、Colligan et al.，eds.，Current Protocolsin Immunology，Greene Publishing Assoc，and Wiley InterScienceN.Y.，(1992，1993)和Muller，Meth.Enzymol.92，589-601(1983)中发现。A CD38BP of the invention may have any suitable affinity and/or strength for one or more epitopes contained at least in part in CD38. Affinity refers to the strength with which CD38BP binds to this epitope. Typically, affinity is determined by the dissociation constant, _Kd , defined as [Ab]×[Ag]/[Ab-Ag], where [Ab-Ag] is the antibody-antigen complex (or _CD38BP -antigen complex), [Ab] is the molar concentration of unbound antibody (or CD38BP), and [Ag] is the molar concentration of unbound antigen. The affinity constant Ka is defined as 1/K _{d .} Suitable methods for determining specificity and affinity by competition can be found, for example, in Harlow et al., Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1988), Colligan et al., eds., Current Protocols in Immunology, Greene Publishing Assoc, and Wiley InterScience N.Y., (1992, 1993) and Muller, Meth. Enzymol. 92, 589-601 (1983).

CD38BP，特别是本发明的抗CD38抗体可具有至少部分含在CD38中的至少一个抗原决定簇的亲和性的范围为10⁴到约10¹⁰M^-1。典型地，此处术语免疫反应是指CD38BP与CD38抗原决定簇以解离常数K_d低于约10^-4M的结合。CD38BPs, particularly anti-CD38 antibodies of the invention, may have an affinity for at least one epitope contained at least in part in CD38 in the range of 10 ⁴ to about 10 ¹⁰ M ⁻¹ . Typically, the term immune response herein refers to the binding of CD38BP to a CD38 epitope with a dissociation constant K _d lower than about 10 ^-4 M.

CD38BP可具有至少与-003和-005和-024相同的针对CD38的亲和性，在某些技术方案中，具有至少与-003和-005和-024相同的亲和性。亲和性可通过任何其它地方所述的本领域中的方法或其已知的等同方法来测定。一个用于测定亲和性的方法的实施例在Scatchard analysis ofMunson & Pollard，Anal.Biochem.107，220(1980)中提供。结合亲和性也可通过平衡方法测定(例如，酶联免疫吸收检测方法(ELISA)或放射免疫检测(RIA)或动力学分析(例如BIACORE^TM分析)。The CD38BP may have at least the same affinity for CD38 as -003 and -005 and -024, and in some embodiments, at least the same affinity as -003 and -005 and -024. Affinity can be determined by methods in the art as described elsewhere or by known equivalent methods. An example of a method for determining affinity is provided in Scatchard analysis of Munson & Pollard, Anal. Biochem. 107, 220 (1980). Binding affinity can also be determined by equilibrium methods (eg, enzyme-linked immunosorbent assay (ELISA) or radioimmunoassay (RIA) or kinetic assays (eg, BIACORE ^™ assay).

典型地，诸如抗CD38抗体这样的本发明的CD38BPs的解离常数小于约100nM、小于约50nM、小于约10nM、小于约5nM或更少、小于约1nM或更少、小于约0.5nM或更少、小于约0.1nM或更少、小于约0.01nM或更少、或甚至小于约0.001nM或更少。Typically, CD38BPs of the invention, such as anti-CD38 antibodies, have a dissociation constant of less than about 100 nM, less than about 50 nM, less than about 10 nM, less than about 5 nM or less, less than about 1 nM or less, less than about 0.5 nM or less , less than about 0.1 nM or less, less than about 0.01 nM or less, or even less than about 0.001 nM or less.

诸如抗CD38抗体这样的本发明的CD38BPs可具有与-003和-005和-024相同的功能特征，例如可通过抗体依赖的细胞毒素作用(ADCC)和补体介导的细胞毒素作用(CDC)检测法进行测定(参见，例如US5500362)。CD38BPs of the invention, such as anti-CD38 antibodies, may have the same functional characteristics as -003 and -005 and -024, e.g. detectable by antibody-dependent cytotoxicity (ADCC) and complement-mediated cytotoxicity (CDC) assay (see, eg, US5500362).

在一个技术方案中，如本发明所述的肽不作为CD38的竞争剂，而作为CD38的拮抗剂。CD38的竞争剂是可激活CD38一个或多个功能的分子。这种功能包括黏附中的受体介导和信号事件和(外部)酶活性。另外，作为外部酶，CD38以NAD⁺底物，形成环化ADP-核糖(cADPR)和ADPR，而且也生成尼克酰胺和尼克酰胺-腺苷二核苷磷酸(NAADP)。已表明cADPR作为Ca²⁺从内质网移动的第二信使。除了通过Ca²⁺的信号传导，CD38信号传导可通过与T和B细胞上的抗原-受体复合物或其它类型的受体复合物，例如MHC分子的交流而发生，并以这种方式参与到几种细胞应答中，而且也转换并分泌IgG1。In a technical solution, the peptide according to the invention does not act as a competitor of CD38, but as an antagonist of CD38. Competitors of CD38 are molecules that activate one or more functions of CD38. Such functions include receptor-mediated and signaling events in adhesion and (external) enzymatic activity. In addition, as an exogenous enzyme, CD38 forms cyclized ADP-ribose (cADPR) and ADPR with NAD ⁺ substrate, and also generates nicotinamide and nicotinamide-adenosine dinucleoside phosphate (NAADP). cADPR has been shown to act as a second messenger for the movement of Ca2 ⁺ from the endoplasmic reticulum. In addition to signaling through Ca2 ⁺ , CD38 signaling can occur through communication with antigen-receptor complexes on T and B cells or other types of receptor complexes, such as MHC molecules, and in this way participate in to several cellular responses and also converts and secretes IgG1.

在一个技术方案中，如本发明所述的肽不诱导PBMCs的显著增殖。在一个技术方案中，如本发明所述的肽不诱导IL-6水平的显著释放。在一个技术方案中，如本发明所述的肽不诱导可检测的IFN-γ水平的释放。这种检测可如Ausiello et al.，Tissue antigens56，538-547(2000)所述进行测定。In a technical solution, the peptides according to the invention do not induce significant proliferation of PBMCs. In one embodiment, the peptides according to the invention do not induce significant release of IL-6 levels. In one embodiment, the peptide according to the invention does not induce the release of detectable levels of IFN-γ. Such detection can be performed as described by Ausiello et al., Tissue antigens 56, 538-547 (2000).

本发明的抗CD38抗体和本发明的其它CD38BPs可通过重组表达在任何合适类型的细胞或动物中产生。Anti-CD38 antibodies of the invention and other CD38BPs of the invention can be produced by recombinant expression in any suitable type of cell or animal.

重组CD38BPs，例如重组抗体，例如重组人抗体，包括通过重组方式制备、表达、产生或分离的CD38BPs，例如抗体，例如人抗体，例如用转染到宿主细胞中的重组表达载体表达的CD38BPs，例如人抗体。Recombinant CD38BPs, such as recombinant antibodies, such as recombinant human antibodies, including CD38BPs prepared, expressed, produced or isolated by recombinant means, such as antibodies, such as human antibodies, such as CD38BPs expressed by recombinant expression vectors transfected into host cells, such as human antibody.

重组抗体，例如重组人抗体也包括从重组的组合的人抗体文库中分离的抗体、从动物，例如转基因动物分离的抗体，或通过任何其它包括将人免疫球蛋白编码核酸序列与其它人免疫球蛋白编码核酸和人免疫球蛋白编码基因外源的核酸序列拼接的方式制备、表达、产生或分离的抗体。典型地，重组人抗体具有来源于人胚系免疫球蛋白序列的可变区和恒定区。但在特定技术方案中，这种重组人抗体进行体外突变(或使用动物转基因的人Ig序列时进行体内体细胞突变)，从而使重组抗体的V_L和V_L区的氨基酸序列是来源于人胚系V_H和V_L序列并与其相关，可以是在体内人抗体胚系库中非天然存在的序列。本发明提供了这两种类型的人抗体。Recombinant antibodies, such as recombinant human antibodies also include antibodies isolated from recombinant combinatorial human antibody libraries, antibodies isolated from animals, such as transgenic animals, or by any other method involving combining human immunoglobulin-encoding nucleic acid sequences with other human immunoglobulin Antibodies prepared, expressed, produced or isolated by splicing protein-encoding nucleic acid and exogenous nucleic acid sequence of human immunoglobulin-encoding gene. Typically, recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in a specific technical scheme, this recombinant human antibody undergoes in vitro mutation (or in vivo somatic mutation when animal transgenic human Ig sequences are used), so that the amino acid sequences of the _VL and _VL regions of the recombinant antibody are derived from human The germline _VH and _VL sequences, and related thereto, may be sequences that do not naturally occur in the human antibody germline repertoire in vivo. The present invention provides both types of human antibodies.

重组蛋白产生的合适方法是本领域已知的，参见例如(Sambrook andRussell(eds.)、Molecular cloning，third edition，2001，Cold Spring HarborLaboratoryPress，Cold Spring Harbor，New York，USA。Suitable methods for recombinant protein production are known in the art, see eg (Sambrook and Russell (eds.), Molecular cloning, third edition, 2001, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA.

同样，抗体产生的合适方法是本领域已知的，包括那些描述在例如Harlow etal.，Antibodies：A Laboratory Manual，Cold Spring HarborLaboratory Press，ColdSpring Harbor，N.Y.，(1988)、Harlow and Lane：Using Antibodies：A LaboratoryManual(Cold Spring Harbor LaboratoryPress(1999))、US 4,376,110和Ausubel etal.，eds.，Current ProtocolsIn Molecular Biology，Greene Publishing Assoc，andWiley InterScienceN.Y.，(1987，1992)中的方法。单克隆抗体可通过首次描述在Kohleretal.，Nature 256，495(1975)中的杂交瘤方法，或通过其它已知的随后开发的方法(参见例如，Goding，Monoclonal Antibodies：Principles and Practice，pp.59-103(AcademicPress，1986))制备。用于产生本发明的抗CD38抗体的产生的杂交瘤也在本发明中提供。这种杂交瘤可通过化学融合、电融合或任何其它合适的技术与任何合适类型的骨髓瘤、异骨髓瘤、淋巴母细胞、浆细胞或其等同的细胞和任何合适类型的抗体表达细胞融合。转化的永生B细胞也可用于有效产生本发明的抗体，因此也在本发明中提供。这种细胞可提供标准技术，例如用Epstein Barr病毒或转化基因的转化来制备(参见，例如″ContinuouslyProliferatingHuman Cell Lines Synthesizing Antibody of PredeterminedSpecificity，″Zurawaki，V.R.et al.，in Monoclonal Antibodies，ed.by KennettR.H.etal.，Plenum Press，N.Y.1980，pp 19-33.)。因此，稳定的连续的和/或永生化抗CD38抗体表达细胞和细胞系是本发明的一个特征。含CD38BP编码或CD38BP片段编码核酸的真核和原核细胞(例如，酵母细胞、连续和/或永生化哺乳动物细胞系(例如，淋巴抗体产生细胞衍生的细胞系)、植物细胞、昆虫细胞和诸如大肠杆菌这样的细菌细胞等)在本发明中提供。表达本发明的抗CD38抗体转基因动物，例如非人灵长类、啮齿类(例如大鼠、豚鼠和田鼠-包括其修饰株，例如联合免疫缺陷鼠(SCID和其它免疫兼容的动物株)、狗等也在本发明中提供。Likewise, suitable methods for antibody production are known in the art, including those described in, e.g., Harlow et al., Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., (1988), Harlow and Lane: Using Antibodies: A Laboratory Manual (Cold Spring Harbor Laboratory Press (1999)), US 4,376,110 and methods in Ausubel et al., eds., Current Protocols In Molecular Biology, Greene Publishing Assoc, and Wiley InterScience N.Y., (1987, 1992). Monoclonal antibodies can be obtained by the hybridoma method first described in Kohler et al., Nature 256, 495 (1975), or by other known and subsequently developed methods (see, e.g., Goding, Monoclonal Antibodies: Principles and Practice, pp. 59 -103 (Academic Press, 1986)) prepared. Hybridomas useful for producing the anti-CD38 antibodies of the present invention are also provided in the present invention. Such hybridomas can be fused by chemical fusion, electrofusion or any other suitable technique with any suitable type of myeloma, heteromyeloma, lymphoblastoid, plasma cell or their equivalent and any suitable type of antibody expressing cell. Transformed immortal B cells can also be used to efficiently produce the antibodies of the invention and are therefore also provided in the present invention. Such cells can be prepared using standard techniques, such as transformation with Epstein Barr virus or transforming genes (see, e.g., "Continuously Proliferating Human Cell Lines Synthesizing Antibody of Predetermined Specificity," Zurawaki, V.R. et al., in Monoclonal Antibodies, ed. by Kennett R. H. et al., Plenum Press, N.Y. 1980, pp 19-33.). Accordingly, stable continuous and/or immortalized anti-CD38 antibody expressing cells and cell lines are a feature of the invention. Eukaryotic and prokaryotic cells (e.g., yeast cells, continuous and/or immortalized mammalian cell lines (e.g., cell lines derived from lymphoid antibody producing cells), plant cells, insect cells and such as Bacterial cells such as Escherichia coli, etc.) are provided in the present invention. Transgenic animals expressing the anti-CD38 antibody of the present invention, such as non-human primates, rodents (such as rats, guinea pigs and voles - including modified strains thereof, such as combined immunodeficiency mice (SCID and other immune-compatible animal strains), dogs etc. are also provided in the present invention.

含编码CD38BPs的外源核酸的重组细胞可通过任何合适的技术(例如，用裸露DNA质粒载体、病毒载体、侵入性细菌细胞载体或其它全细胞载体等转染/转化，包括通过磷酸钙沉淀的转染、受体介导的定位和转染、生物弹射击递送、电穿孔、葡聚糖介导的转染、脂质体介导的转化、原生质体融合、直接微注射等方法将CD38BP编码序列(或序列)递送到细胞中)制备。转化/转染细胞的方法是本领域已知的(参见，例如Sambrook et al.，MolecularCloning：A Laboratory Manual，ColdSpring Harbor Laboratory Press(2d Edition，1989 and 3rd Edition，2001)和F.Ausubel et al.，ed.Current Protocols inMolecular Biology，GreenePublishing and Wiley InterScience New York(1987)。这种重组细胞是本发明的一个特征。Recombinant cells containing exogenous nucleic acids encoding CD38BPs can be transfected/transformed by any suitable technique (e.g., with naked DNA plasmid vectors, viral vectors, invasive bacterial cell vectors, or other whole-cell vectors, including by calcium phosphate precipitation). Transfection, receptor-mediated localization and transfection, biolistic delivery, electroporation, dextran-mediated transfection, liposome-mediated transformation, protoplast fusion, direct microinjection and other methods to encode CD38BP Sequence (or sequence) delivery into cells) preparation. Methods of transforming/transfecting cells are known in the art (see, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press (2d Edition, 1989 and 3rd Edition, 2001) and F. Ausubel et al. , ed. Current Protocols in Molecular Biology, Greene Publishing and Wiley InterScience New York (1987). Such recombinant cells are a feature of the present invention.

作为宿主用于重组蛋白蛋白的细胞系是本领域周知的，包括多种可从美国典型培养物保藏中心(ATCC)获得的永生化细胞系。这些细胞系中包括、中国仓鼠卵巢(CHO)细胞、NSO、SP2细胞、HeLa细胞、幼仓鼠肾(BHK)细胞、猴肾细胞(COS)、人肝癌细胞(例如Hep G2)、A549细胞及多种其它细胞系。其它可用的细胞系是诸如Sf9细胞这样的昆虫细胞系。当编码诸如CD38BPs(包括抗CD38抗体)这样的蛋白的核酸(或含核酸的载体)导入到哺乳动物宿主细胞中时，诸如CD38BPs这样的蛋白可通过将宿主细胞培养足以在宿主细胞中表达诸如CD38BPs这样的蛋白或将诸如CD38BPs这样的蛋白分泌到宿主细胞生长的培养基中的时间而产生。可用标准蛋白纯化方法从培养基中回收CD38BPs。当没有分泌信号而直接表达时，也可从宿主细胞裂解物中回收CD38BPs。Cell lines for use as hosts for recombinant proteins are well known in the art and include a variety of immortalized cell lines available from the American Type Culture Collection (ATCC). These cell lines include, Chinese hamster ovary (CHO) cells, NSO, SP2 cells, HeLa cells, baby hamster kidney (BHK) cells, monkey kidney cells (COS), human liver cancer cells (e.g. Hep G2), A549 cells and many more. other cell lines. Other useful cell lines are insect cell lines such as Sf9 cells. When a nucleic acid (or a nucleic acid-containing vector) encoding a protein such as CD38BPs (including an anti-CD38 antibody) is introduced into a mammalian host cell, the protein such as CD38BPs can be expressed in the host cell sufficiently by culturing the host cell such as CD38BPs Such proteins are produced when proteins such as CD38BPs are secreted into the medium in which the host cells are grown. CD38BPs can be recovered from the culture medium using standard protein purification methods. CD38BPs can also be recovered from host cell lysates when expressed directly without a secretion signal.

CD38BPs，例如抗CD38抗体，也可在细菌细胞和真核单细胞微生物，例如酵母中产生。产生诸如抗CD38抗体这样的CD38BPs的细菌细胞典型地缺少正常的糖基化，因此细菌细胞产生的抗CD38抗体或多或少会在哺乳动物细胞和/或动物中产生的抗CD38抗体相关的ADCC功能和其它免疫应答方面(例如，NK细胞的募集)有缺陷。酵母细胞产生的CD38BPs，例如抗CD38抗体通常具有与哺乳动物细胞中产生的抗体类型不同的糖基化模式。但在酵母中产生具有有效糖基化的抗体的方法目前已由公司，例如Glycofi，Inc.(Lebanon，NH，USA)开发出来。也参见Wildt S et al.，Nat Rev Microbiol.3(2)，119-28(2005)。CD38BPs, such as anti-CD38 antibodies, can also be produced in bacterial cells and eukaryotic unicellular microorganisms, such as yeast. Bacterial cells that produce CD38BPs such as anti-CD38 antibodies typically lack normal glycosylation, so anti-CD38 antibodies produced by bacterial cells will more or less ADCC associated with anti-CD38 antibodies produced in mammalian cells and/or animals Functional and other aspects of the immune response (eg, recruitment of NK cells) are defective. CD38BPs produced in yeast cells, such as anti-CD38 antibodies, often have different glycosylation patterns than those produced in mammalian cells. However, methods for producing antibodies with efficient glycosylation in yeast have now been developed by companies such as Glycofi, Inc. (Lebanon, NH, USA). See also Wildt S et al., Nat Rev Microbiol. 3(2), 119-28 (2005).

当编码CD38BP基因(包括抗CD38抗体基因)的重组表达载体导入到哺乳动物宿主细胞中时，可通过将宿主细胞培养足以在宿主细胞中表达CD38BP或将抗体分泌到宿主细胞生长的培养基中的时间而产生CD38BPs。抗体和其它CD38BPs从细胞培养物、细胞裂解物和动物(例如，从产生抗CD38抗体的转基因动物的腹水液中)中的纯化可通过使用任何本领域已知的合适技术，例如免疫亲和柱纯化；硫酸盐沉淀；色谱聚焦；制备型SDS-PAGE等来完成。When a recombinant expression vector encoding a CD38BP gene (including an anti-CD38 antibody gene) is introduced into a mammalian host cell, it can be obtained by culturing the host cell enough to express CD38BP in the host cell or secrete the antibody into the growth medium of the host cell time to produce CD38BPs. Antibodies and other CD38BPs can be purified from cell cultures, cell lysates, and animals (e.g., from ascites fluid of transgenic animals producing anti-CD38 antibodies) by using any suitable technique known in the art, such as immunoaffinity columns Purification; sulfate precipitation; chromatographic focusing; preparative SDS-PAGE, etc. to complete.

本发明的人单克隆抗体也可通过各种其它技术，包括传统的单克隆抗体方法，例如Kohler and Milstein，Nature 256，495(1975)中所述的标准体细胞杂交技术来产生。其它产生单克隆抗体的技术也可使用，例如使用人抗体基因文库的噬菌体展示技术。在一个技术方案中，通过使用杂交瘤产生的本发明的抗CD38抗体在鼠系统中产生。在鼠中产生杂交瘤是非常成熟的流程。免疫流程和分离用于融合的免疫的脾细胞的技术是本领域已知的。融合部分(例如鼠骨髓瘤细胞)和融合程序也是已知的。Human monoclonal antibodies of the invention can also be produced by a variety of other techniques, including traditional monoclonal antibody methods, eg, standard somatic cell hybridization techniques as described in Kohler and Milstein, Nature 256, 495 (1975). Other techniques for producing monoclonal antibodies can also be used, such as phage display using human antibody gene libraries. In one embodiment, the anti-CD38 antibodies of the invention produced by using hybridomas are produced in a murine system. Generation of hybridomas in mice is a well-established procedure. Immunization protocols and techniques for isolating immunized splenocytes for fusion are known in the art. Fusion moieties (eg, murine myeloma cells) and fusion procedures are also known.

为了产生全长的针对CD38的人单克隆抗体，含人免疫球蛋白基因的转基因或转染色体鼠(例如HCo12、HCo7或KM鼠)可用CD38抗原和/或表达CD38的细胞的富集产物来免疫，例如，如Lonberg et al.，(1994)、同上，Fishwild et al.，(1996)、同上，及WO 98/24884所述。选择性地，可用编码人CD38的DNA免疫鼠。在第一次融合时鼠可以为6-16周龄。例如，CD38抗原的富集产物(5-50μg)可用于腹膜内免疫HuMAb鼠。在免疫过程中使用纯化的或富集的CD38抗原产物未产生抗体时，也可以用表达CD38的细胞，例如细胞系来免疫鼠，以促进免疫应答。To generate full-length human monoclonal antibodies directed against CD38, transgenic or transchromosomal mice containing human immunoglobulin genes (eg, HCo12, HCo7, or KM mice) can be immunized with CD38 antigen and/or enriched products of CD38-expressing cells , for example, as described in Lonberg et al., (1994), supra, Fishwild et al., (1996), supra, and WO 98/24884. Alternatively, mice can be immunized with DNA encoding human CD38. Mice can be 6-16 weeks old at first fusion. For example, an enriched product (5-50 μg) for CD38 antigen can be used to immunize HuMAb mice intraperitoneally. When antibodies are not produced during immunization using purified or enriched CD38 antigen products, CD38-expressing cells, such as cell lines, can also be used to immunize mice to promote an immune response.

使用各种抗原的积累经验表明，当用含在完全弗氏佐剂中的CD38表达细胞进行腹膜内(i.p.)或皮下(s.c.)初始免疫，然后每隔1周用含在PBS中的CD38表达细胞进行i.p.免疫(共10次以上)，可以在HuMAb转基因鼠中产生最大的应答。可在免疫流程中用眶后采血获得的血浆样品来监测免疫应答。可通过FACS分析筛选血浆，具有足够滴度的抗CD38人免疫球蛋白的鼠可用于融合。鼠可在处死并除去脾的3天前用CD38表达细胞进行静脉内注射，如实施例4所述。Accumulated experience with various antigens shows that when priming intraperitoneally (i.p.) or subcutaneously (s.c.) with CD38-expressing cells in complete Freund's adjuvant, followed by CD38-expressing cells in PBS every other week Cells were i.p. immunized (more than 10 times in total) to generate the greatest response in HuMAb transgenic mice. Immune responses can be monitored during the immunization protocol using plasma samples obtained by retro-orbital bleeding. Plasma can be screened by FACS analysis and mice with sufficient titers of anti-CD38 human immunoglobulin can be used for fusions. Mice can be injected intravenously with CD38 expressing cells, as described in Example 4, 3 days before sacrifice and removal of the spleen.

为了产生可生产针对人CD38的人单克隆抗体的杂交瘤，从免疫鼠中分离脾细胞和淋巴结细胞，并与合适的永生化细胞系，例如鼠骨髓瘤细胞系融合。然后筛选可产生抗原特异的抗体的杂交瘤。例如，将来自免疫鼠的脾淋巴细胞的单细胞悬液与SP2/0非分泌鼠骨髓瘤细胞(ATCC，CRL 1581)用50％PEG(w/v)融合。在平底微孔板中放置细胞，每孔约1×10⁵，然后在除通用试剂外，含10％胎克隆血清、5-10％Origen杂交瘤克隆因子(IGEN)1×HAT(Sigma)的选择培养基中培养2周。约2周后，可在用HT替换HAT的培养基中培养细胞。然后通过针对含κ-轻链的抗体的ELISA和通过使用具有CD38特异性的CD38表达细胞的FACS分析来筛选各个孔。当大量的杂交瘤生长时，在10-14天后观察培养基。可重新放置分泌抗体的杂交瘤，再次筛选，如果对人IgG仍为阳性，则可将抗CD38单克隆抗体通过有限稀释至少亚克隆2次。然后在体外培养稳定的亚克隆，以在组织培养基中产生用于鉴定的抗体。To generate hybridomas producing human monoclonal antibodies directed against human CD38, splenocytes and lymph node cells are isolated from immunized mice and fused with a suitable immortalized cell line, such as a murine myeloma cell line. Hybridomas are then screened for the production of antigen-specific antibodies. For example, single cell suspensions of splenic lymphocytes from immunized mice were fused with SP2/0 non-secreting murine myeloma cells (ATCC, CRL 1581) with 50% PEG (w/v). Place the cells in a flat-bottomed microplate, about 1×10 ⁵ per well, and then in addition to the common reagents, contain 10% fetal clone serum, 5-10% Origen hybridoma cloning factor (IGEN) 1×HAT (Sigma) Cultured in selective medium for 2 weeks. After about 2 weeks, the cells can be cultured in medium replacing HAT with HT. Individual wells were then screened by ELISA against kappa-light chain containing antibodies and by FACS analysis using CD38 expressing cells with CD38 specificity. The medium was observed after 10-14 days when a large number of hybridomas had grown. Antibody-secreting hybridomas can be relocated and screened again, and if they are still positive for human IgG, anti-CD38 monoclonal antibodies can be subcloned at least twice by limiting dilution. Stable subclones are then cultured in vitro to produce antibodies for characterization in tissue culture medium.

本发明的人抗体也可在宿主细胞转染瘤中联合使用例如本领域已知的重组DNA技术和基因转染方法来产生，参见例如Morrison，S.， Science 229，1202(1985)。Human antibodies of the invention can also be produced in host cell transfectomas using, for example, combinations of recombinant DNA techniques and gene transfection methods known in the art, see, eg, Morrison, S., Science 229, 1202 (1985).

例如，为了表达抗体或其抗体片段，可通过标准分子生物学技术(例如PCR、定点突变)获得编码部分或全长轻链和重链的DNAs，并将其插入到表达载体中，从而使基因与转录和翻译控制序列可操作连接。在本文中，术语“可操作连接”是指抗体基因与载体连接，从而使载体中的转录和翻译控制序列行使调控抗体基因的转录和翻译的功能。表达载体和表达控制序列选择为与所用的表达宿主细胞兼容的序列。可将抗体轻链基因和抗体重链基因插入到不同载体中，或更典型地，两个基因插入到相同的表达载体中。可通过标准方法(例如，抗体基因片段和载体的互补限制性位点的连接，或如果不存在限制性位点，则进行平末端连接)将抗体基因插入到表达载体中。此处所述的抗体轻链和重链可变区可通过将任何抗体同种型的全长抗体基因插入到已有编码所需同种型的重链恒定区和轻链恒定区的表达载体中，从而使V_H片段与载体中的C_H片段可操作连接，V_L片段与载体中的C_L片段可操作连接而获得。另外，或选择性地，重组表达载体可编码利于抗体链从宿主细胞中分泌的信号肽。可将抗体链基因克隆到载体中，从而使信号肽同框连接到抗体链基因的氨基末端。信号肽可以是免疫球蛋白的信号肽或异源信号肽(例如，来自非免疫球蛋白的蛋白信号肽)。For example, to express an antibody or antibody fragment thereof, DNAs encoding partial or full-length light and heavy chains can be obtained by standard molecular biology techniques (e.g., PCR, site-directed mutagenesis) and inserted into expression vectors so that the genes Operably linked to transcriptional and translational control sequences. Herein, the term "operably linked" means that the antibody gene is linked to the carrier, so that the transcription and translation control sequences in the carrier can perform the function of regulating the transcription and translation of the antibody gene. The expression vector and expression control sequences are selected to be those compatible with the expression host cell used. The antibody light chain gene and the antibody heavy chain gene can be inserted into separate vectors or, more typically, both genes are inserted into the same expression vector. Antibody genes can be inserted into expression vectors by standard methods (eg, ligation of antibody gene fragments and complementary restriction sites of the vector, or blunt end ligation if no restriction sites are present). The antibody light and heavy chain variable regions described herein can be obtained by inserting the full-length antibody gene of any antibody isotype into an existing expression vector encoding the heavy and light chain constant regions of the desired isotype , so that the _VH segment is operably linked to the _CH segment in the vector, and the _VL segment is operably linked to the _CL segment in the vector. Additionally, or alternatively, the recombinant expression vector may encode a signal peptide that facilitates secretion of the antibody chain from the host cell. The antibody chain genes can be cloned into a vector such that the signal peptide is linked in-frame to the amino terminus of the antibody chain genes. The signal peptide can be that of an immunoglobulin or a heterologous signal peptide (eg, a signal peptide from a protein other than an immunoglobulin).

除了抗体链基因，本发明的重组表达载体可携带调控抗体链基因在宿主细胞中表达的调控序列。In addition to antibody chain genes, the recombinant expression vector of the present invention can carry regulatory sequences that regulate the expression of antibody chain genes in host cells.

除了抗体链基因和调控序列外，本发明的重组表达载体可携带其它序列，例如调控载体在宿主细胞中复制的序列(例如复制起点)和选择标记基因。选择标记基因利于在引入载体的宿主细胞中进行筛选(参见例如US 4,399,216、US 4,634,665和US 5,179,017)。例如，典型地，选择标记基因可在引入载体的宿主细胞中赋予对药物，例如G418、潮霉素或甲氨蝶呤的抗性。选择标记基因的例子包括二氢叶酸还原酶(DHFR)基因(用于在dhfr宿主细胞中用甲氨蝶呤筛选/扩增)和neo基因(用于G418筛选)。In addition to antibody chain genes and regulatory sequences, the recombinant expression vectors of the present invention may carry other sequences, such as sequences that regulate replication of the vector in host cells (eg, origin of replication) and selectable marker genes. A selectable marker gene facilitates selection in the host cell into which the vector is introduced (see eg US 4,399,216, US 4,634,665 and US 5,179,017). For example, a selectable marker gene typically confers resistance to a drug, such as G418, hygromycin, or methotrexate, in the host cell into which the vector is introduced. Examples of selectable marker genes include the dihydrofolate reductase (DHFR) gene (for selection/amplification with methotrexate in dhfr host cells) and the neo gene (for G418 selection).

为了表达轻链和重链，将编码重链和轻链的表达载体通过标准技术转染到宿主细胞中。宿主细胞可以是原核或真核宿主细胞，例如哺乳动物。例如，可在原核宿主细胞中表达抗原结合片段，在真核宿主细胞中表达全长抗体。To express the light and heavy chains, expression vectors encoding the heavy and light chains are transfected into host cells by standard techniques. The host cell can be a prokaryotic or eukaryotic host cell, such as a mammal. For example, antigen-binding fragments can be expressed in prokaryotic host cells and full-length antibodies can be expressed in eukaryotic host cells.

在一个技术方案中，抗体在真核细胞，例如哺乳动物宿主细胞中表达。在哺乳动物宿主细胞中表达本发明的重组抗体的实施例包括CHO细胞(包括dhfr-CHO细胞，在Urlauband Chasin，PNAS USA 77，4216-4220(1980)描述，以DHFR为筛选标记，例如在R.J.KaufmanandP.A.Sharp，MoI.Biol.159，601-621(1982)中描述)、NS/O骨髓瘤细胞、COS细胞、HEK293细胞和SP2.0细胞。特别是使用NS/O骨髓瘤细胞，表达系统的另一个实施例是GS(谷氨酸合成酶)基因表达系统，在WO87/04462、WO89/01036和EP338841中描述。In one embodiment, antibodies are expressed in eukaryotic cells, such as mammalian host cells. Examples of expressing the recombinant antibody of the present invention in mammalian host cells include CHO cells (including dhfr-CHO cells, described in Urlauband Chasin, PNAS USA 77, 4216-4220 (1980), using DHFR as a selection marker, such as in R.J. Kaufman and P.A. Sharp, MoI. Biol. 159, 601-621 (1982) described), NS/O myeloma cells, COS cells, HEK293 cells and SP2.0 cells. Another example of an expression system, particularly using NS/O myeloma cells, is the GS (glutamate synthetase) gene expression system described in WO87/04462, WO89/01036 and EP338841.

可在其它表达系统，包括原核细胞，例如微生物，例如产生scFv抗体的大肠杆菌、藻类和昆虫细胞中表达CD38BP。另外，可在转基因非人动物，例如羊和兔奶或鸡蛋，或转基因植物中表达CD38BPs。参见例如Verma，R.et al.，J.lmmunol.Meth.216，165-181(1998)、Pollocket al.，J.lmmunol.Meth.23J.，147-157(1999)和Fischer，R.et al.，Biol.Chem.380，825-839(1999)。CD38BP can be expressed in other expression systems, including prokaryotic cells, such as microorganisms such as E. coli, algae and insect cells that produce scFv antibodies. Additionally, CD38BPs can be expressed in transgenic non-human animals, such as sheep and rabbit milk or eggs, or in transgenic plants. See eg Verma, R. et al., J. Immunol. Meth. 216, 165-181 (1998), Pollock et al., J. Immunol. Meth. 23 J., 147-157 (1999) and Fischer, R. et al., Biol. Chem. 380, 825-839 (1999).

本发明的双特异性和多特异性CD38BPs可通过化学技术、“Polydoma”技术(参见US4,474,893)或重组DNA技术制备(参见例如D.M.Kranz et al.，PNAS USA 78，5807(1981))。The bispecific and multispecific CD38BPs of the invention can be prepared by chemical techniques, "Polydoma" techniques (see US 4,474,893) or recombinant DNA techniques (see eg D.M. Kranz et al., PNAS USA 78, 5807 (1981)).

本发明的双特异性抗体可通过各种已知的方法，包括杂交瘤融合或Fab′片段连接(参见，例如Songsivilai&Lachmann，Clin.Exp.Immunol.79，315-321(1990)和Kostelny etal.，J.Immunol.M8，1547-1553(1992))来产生。传统地，双特异性抗体的重组产生是基于两个免疫球蛋白重链-轻链对的共表达，其中两个重链具有不同的特异性(参见例如，Milsteinand Cuello，Nature 305，537(1983))。由于免疫球蛋白重链和轻链的随机分选，这些杂交瘤(四源杂交瘤)可产生约10中不同抗体分子的混合物，其中只有一种具有正确的双特异性结构。类似流程在WO93/08829和Traunecker et al.，EMBO J.10，3655(1991)中描述。The bispecific antibody of the present invention can be obtained by various known methods, including fusion of hybridomas or linking of Fab' fragments (see, for example, Songsivilai & Lachmann, Clin. Exp. Immunol. 79, 315-321 (1990) and Kostelny et al., J. Immunol. M8, 1547-1553 (1992)) to produce. Traditionally, the recombinant production of bispecific antibodies is based on the co-expression of two immunoglobulin heavy chain-light chain pairs, where the two heavy chains have different specificities (see, e.g., Milstein and Cuello, Nature 305, 537 (1983 )). Due to the random sorting of immunoglobulin heavy and light chains, these hybridomas (quadromas) produce a mixture of about 10 different antibody molecules, only one of which has the correct bispecific structure. Similar procedures are described in WO93/08829 and Traunecker et al., EMBO J. 10, 3655 (1991).

根据不同的方法，将具有所需结合特异性的抗体可变结构域(抗体-抗原结合位点)与免疫球蛋白恒定结构域序列通过重组或合成方法融合在一起。可变结构域序列典型地与含至少部分铰链、C_H2和C_H3区的免疫球蛋白重链恒定结构域融合。而且典型地，在融合肽中至少也存在一个含轻链结合所需位点的第一重链恒定区(C_H1)。在这种方法的具体实施例中，产生的双特异性抗体含有在一个臂具有第一结合特异性的免 疫球蛋白重链的杂交体，另一个臂为免疫球蛋白重链-轻链对(提供第二结合特异性)的杂交体。这种非对称结构可利于从不需要的免疫球蛋白链组合中分离所需的双特异性化合物(这种方法描述在WO94/04690中)。对产生双特异性抗体的进一步细节，参见例如Suresh et al.，MethodsinEnzymology 121，210(1986)。According to different approaches, antibody variable domains (antibody-antigen combining sites) with the desired binding specificities are fused to immunoglobulin constant domain sequences by recombinant or synthetic methods. The variable domain sequences are typically fused to an immunoglobulin heavy chain constant domain comprising at least part of the hinge, _{CH2 and CH3 regions} . And typically, at least one first heavy chain constant region ( _CH1 ) containing the site required for light chain binding is also present in the fusion peptide. In specific embodiments of this method, bispecific antibodies are produced that contain a hybrid of an immunoglobulin heavy chain with a first binding specificity in one arm and an immunoglobulin heavy chain-light chain pair ( providing a hybrid of the second binding specificity). This asymmetric structure may facilitate the separation of desired bispecific compounds from unwanted combinations of immunoglobulin chains (this approach is described in WO94/04690). For further details on the production of bispecific antibodies see, eg, Suresh et al., Methods in Enzymology 121, 210 (1986).

在另一种方法中，一对抗体分子的交界处通过构建以使从重组细胞培养物中回收的异源二聚体，从而形成双特异性抗体分子群的比例最大化。典型地，这种交界处含至少一部分抗体恒定区的C_H3结构域。通常在这种方法中，将第一抗体分子交界处的一个和多个具有较小的侧链的氨基酸残基用具有较大侧链的氨基酸残基(例如酪氨酸或色氨酸)替代。通过用较小残基(例如丙氨酸或苏氨酸)替代较大氨基酸侧链残基，则在第二抗体分子交界处产生与大侧链氨基酸残基具有相同或相似大小的补偿性“洞穴”。这可增加异源二聚体而不是其它不需要的诸如同源二聚体这样的终产物的产量。In another approach, the junction of a pair of antibody molecules is engineered to maximize the proportion of heterodimers recovered from recombinant cell culture, thereby forming the population of bispecific antibody molecules. Typically, this junction comprises at least a portion of the _CH3 domain of an antibody constant region. Typically in this approach, one or more amino acid residues with smaller side chains at the junction of the first antibody molecule are replaced with amino acid residues with larger side chains (such as tyrosine or tryptophan) . By substituting smaller residues (such as alanine or threonine) for larger amino acid side chain residues, a compensatory "compensatory" amino acid residue of the same or similar size as the large side chain amino acid residue is created at the junction of the second antibody molecule cave". This increases the yield of heterodimers rather than other unwanted end products such as homodimers.

本发明的双特异性和多特异性分子可通过本领域已知的方法偶联具有结合特异性，例如抗FcR和抗CD38结合特异性的构建体而制备。当结合特异性是蛋白或肽时，可使用各种偶联或交联试剂进行共价偶联。交联试剂的实施例包括蛋白A、碳二亚胺、S-乙酰基-巯基乙酸N-羟基丁二酰亚胺酯(SATA)、5，5′双(4-硝基-3-羧基苯)二硫化物(DTNB)、o-亚苯基顺丁烯二酰亚胺(oPDM)、3-(2-吡啶二巯基)丙酸N-羟基琥珀酰亚胺酯(SPDP)和磺基琥珀酰亚胺基4-[马来酰亚胺甲基]-环己烷-1羧酸盐(sulfo-SMCC)，参见，例如Karpovsky etal.，J.Exp.Med.160，1686(1984)、Liu，M.A.et al.，PNAS USA 82，8648(1985)。在另一个实施例中，Brennan et al.，Science 229，81(1985)描述了将完整抗体蛋白酶裂解产生F(ab′)₂片段的流程。这些片段在存在可稳定邻近二硫，并抑制分子内二硫键形成的巯醇配位试剂亚砷酸钠时被还原。然后将产生的Fab′片段转变为硝基苯甲酸(TNB)衍生物。然后将其中一种Fab′-TNB衍生物通过用半胱胺还原再次转变为Fab′-巯醇，并与等量的其它Fab′-TNB衍生物混合而形成双特异性抗体。Shalabyet al.，J.Exp.Med.175，217-225(1992)根据相关技术，描述了完全人源化双特异性抗体F(ab′)₂分子的产生。其它方法包括那些Paulus(Behring Ins.Mitt.No.78，118-132(1985))和Glennie et al.，J.Immunol. 139，2367-2375(1987)描述的方法。偶联试剂的实施例有SATA和sulfo-SMCC，均可从Pierce ChemicalCo.(Rockford，IL)获得。Bispecific and multispecific molecules of the invention can be prepared by conjugating constructs with binding specificities, such as anti-FcR and anti-CD38 binding specificities, by methods known in the art. When the binding specificity is a protein or peptide, various coupling or cross-linking reagents can be used for covalent coupling. Examples of crosslinking reagents include protein A, carbodiimide, N-hydroxysuccinimide S-acetyl-thioglycolate (SATA), 5,5'bis(4-nitro-3-carboxyphenyl ) disulfide (DTNB), o-phenylenemaleimide (oPDM), 3-(2-pyridyldimercapto)propionic acid N-hydroxysuccinimide ester (SPDP) and sulfosuccinate Imido 4-[maleimidomethyl]-cyclohexane-1 carboxylate (sulfo-SMCC), see, for example, Karpovsky et al., J. Exp. Med. 160, 1686 (1984), Liu, MA et al., PNAS USA 82, 8648 (1985). In another example, Brennan et al., Science 229, 81 (1985) describe a procedure for the proteolytic cleavage of intact antibodies to generate F(ab') ₂ fragments. These fragments are reduced in the presence of the thiol complexing reagent sodium arsenite, which stabilizes adjacent disulfides and inhibits intramolecular disulfide bond formation. The resulting Fab' fragments are then converted to nitrobenzoic acid (TNB) derivatives. One of the Fab'-TNB derivatives was then reconverted to Fab'-thiol by reduction with cysteamine and mixed with an equal amount of the other Fab'-TNB derivative to form a bispecific antibody. Shalaby et al., J. Exp. Med. 175, 217-225 (1992) describe the generation of fully humanized bispecific antibody F(ab') ₂ molecules according to the related art. Other methods include those described by Paulus (Behring Ins. Mitt. No. 78, 118-132 (1985)) and Glennie et al., J. Immunol. 139, 2367-2375 (1987). Examples of coupling reagents are SATA and sulfo-SMCC, both available from Pierce Chemical Co. (Rockford, IL).

当结合特异性是抗体时，它们可通过两个重链铰链区的C末端巯基键偶联。在一个技术方案中，可在偶联前修饰铰链区使其含奇数数目例如1个的巯基残基。When the binding specificities are antibodies, they can be coupled via the C-terminal sulfhydryl bonds of the hinge regions of the two heavy chains. In one embodiment, the hinge region can be modified to contain an odd number, eg, 1, of sulfhydryl residues prior to conjugation.

选择性地，两种结合特异性可在同一个载体上编码，并在同一个宿主细胞中表达和组装。这种方法在双特异性和多特异性分子是mAb×mAb、mAb×Fab、Fab×F(ab′)₂或配体×Fab融合蛋白时特别有用。本发明的双特异性和多特异性分子，例如双特异性分子可以是单链分子，例如单链双特异性抗体、含一个单链抗体和结合决定簇的双特异性分子，或含两个结合决定簇的单链双特异性分子。双特异性和多特异性分子也可以是单链分子或可含有至少两个单链分子。制备双和多特异性分子的方法在例如US 5,260,203、US 5,455,030、US4,881,175、US5,132,405、US 5,091,513、US 5,476,786、US 5,013,653、US 5,258,498和US5,482,858中描述。Alternatively, both binding specificities can be encoded on the same vector, expressed and assembled in the same host cell. This approach is particularly useful when the bispecific and multispecific molecules are mAbxmAb, mAbxFab, FabxF(ab') ₂ or LigandxFab fusion proteins. The bispecific and multispecific molecules of the invention, e.g., bispecific molecules, can be single chain molecules, e.g., single chain bispecific antibodies, bispecific molecules comprising a single chain antibody and a binding determinant, or comprising two Single-chain bispecific molecules that bind determinants. Bispecific and multispecific molecules can also be single-chain molecules or can contain at least two single-chain molecules. Methods of making bi- and multispecific molecules are described, for example, in US 5,260,203, US 5,455,030, US 4,881,175, US 5,132,405, US 5,091,513, US 5,476,786, US 5,013,653, US 5,258,498 and US 5,482,858.

已描述了各种直接从重组细胞培养物中制备和分离双特异性抗体片段的技术。例如，可通过亮氨酸拉链产生双特异性抗体(参见，例如Kostelny et al.，J.Immunol.148(5)，1547-1553(1992))。来自Fos和Jun蛋白的亮氨酸拉链肽可与两个不同抗体的Fab′部分通过基因融合而连接，得到的抗体同源二聚体在铰链区还原，从而形成可重新氧化的单体，以形成抗体异源二聚体。在中Hollinger et al.，PNAS USA 90，6444-6448(1993)描述的“双功能抗体”技术也提供了制备双特异性抗体片段的选择方案。已报道另一种使用单链Fv(sFv)二聚体制备双特异性抗体片段的策略是。参见例如Gruber et al.，J.Immunol.152，5368(1994)。Various techniques have been described for the preparation and isolation of bispecific antibody fragments directly from recombinant cell culture. For example, bispecific antibodies can be generated via leucine zippers (see, eg, Kostelny et al., J. Immunol. 148(5), 1547-1553 (1992)). Leucine zipper peptides from the Fos and Jun proteins can be linked by gene fusion to the Fab' portions of two different antibodies, and the resulting antibody homodimers are reduced at the hinge region to form reoxidizable monomers to Formation of antibody heterodimers. The "bifunctional antibody" technique described in Hollinger et al., PNAS USA 90, 6444-6448 (1993) also provides an option for preparing bispecific antibody fragments. Another strategy for preparing bispecific antibody fragments using single-chain Fv (sFv) dimers has been reported. See eg Gruber et al., J. Immunol. 152, 5368 (1994).

另外，双特异性抗体可形成“双功能抗体”(Holliger et al.，PNASUSA，90，6444-6448(1993))或“Janusins”(Traunecker et al.，EMBOJ10，3655-3659(1991)和Trauneckeret al.，lnt J Cancer Suppl 7，51-52(1992))。定义的双特异性抗体不存在具有单个结合位点的片段形式(例如Fab、Fab′和Fv片段，也由本发明提供)。Alternatively, bispecific antibodies can form "diabodies" (Holliger et al., PNASUSA, 90, 6444-6448 (1993)) or "Janusins" (Traunecker et al., EMBOJ10, 3655-3659 (1991) and Trauneckeret al., Int J Cancer Suppl 7, 51-52 (1992)). A defined bispecific antibody does not exist in fragment form with a single binding site (eg Fab, Fab' and Fv fragments, also provided by the present invention).

双特异性和多特异性分子与其靶标的结合可通过酶联免疫吸收测定法(ELISA)、放射性免疫检测(RIA)、FACS分析、生物检测(例 如生长抑制)或Western印迹检测法来确定。这些方法每种通常都是通过使用针对目的复合物的标记试剂(例如抗体)检测目的蛋白-抗体复合物的存在。例如，FcR-抗体复合物可用例如识别和特异结合抗体-FcR复合物的酶联抗体或抗体片段来检测。选择性地，可通过任何其它各种免疫检测法来检测复合物。例如，抗体可进行放射性标记，使用放射性免疫检测法(RIA)来检测(参见，例如Weintraub，B.，Principles ofRadioimmunoassays，Seventh Training Course on RadioligandAssayTechniques，The Endocrine Society，March，1986)。在这种方法中，使用γ计数器或闪烁计数器或通过自显影来检测放射性同位素。Binding of bispecific and multispecific molecules to their targets can be determined by enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), FACS analysis, biological assays (e.g., growth inhibition), or Western blot assays. Each of these methods typically detects the presence of a protein-antibody complex of interest by using a labeled reagent (eg, an antibody) directed against the complex of interest. For example, an FcR-antibody complex can be detected using, for example, an enzyme-linked antibody or antibody fragment that recognizes and specifically binds to the antibody-FcR complex. Alternatively, the complex can be detected by any other various immunoassays. For example, antibodies can be radiolabeled and detected using radioimmunoassays (RIA) (see, e.g., Weintraub, B., Principles of Radioimmunoassays, Seventh Training Course on Radiolig and Assay Techniques, The Endocrine Society, March, 1986). In this method, radioisotopes are detected using a gamma counter or scintillation counter or by autoradiography.

如前所述，抗体主要通过位于六个重链和轻链互补决定区(CDRs)的氨基酸残基与靶抗原相互作用。本发明提供具有与来源于-003或-005或-024的CDR区相同或其衍生的CDR区的抗体。这种抗体可通过构建包括将来自-003或-005或-024的CDR序列移植到具有不同特性的不同抗体的框架序列中的表达载体而产生。As mentioned earlier, antibodies interact with target antigens primarily through amino acid residues located in the six heavy and light chain complementarity determining regions (CDRs). The present invention provides an antibody having a CDR region identical to or derived from a CDR region derived from -003 or -005 or -024. Such antibodies can be produced by constructing expression vectors comprising grafting of CDR sequences from -003 or -005 or -024 into the framework sequences of different antibodies with different properties.

这种框架序列可从含胚系抗体基因序列的公共DNA数据库获得。这些胚系序列由于不完全包括在B细胞成熟时通过V(D)J连接形成的组装可变基因，因此不同于成熟抗体基因序列。胚系基因序列也不同于在可变基因，典型地集中在CDRs中的高亲和力的第二抗体的序列。例如，体细胞突变在框架区1的氨基末端部分和框架区4的羧基末端部分相对较少。因此，不需获得特定抗体完整的DNA序列来重新产生完整的具有与原始抗体相似结合化学的重组抗体(参见WO 99/45962)。部分跨越CDR区的重链和轻链序列用于测定参与重组抗体可变基因中的胚系可变和连接基因片段。然后用胚系序列填充可变区的缺失部分。重链和轻链引导序列在蛋白成熟时被切割，不参与最终抗体的特性。为了增加缺失的序列，将克隆的cDNA序列与合成的寡核苷酸通过连接或PCR扩增进行连接。选择性地，完整的可变区可通过合成为一组短的重叠的寡核苷酸，并通过PCR合并而产生完整的合成的可变区克隆。该过程具有特定的优势，例如可删除或插入特定的限制性位点，或优化特定的密码子。Such framework sequences are available from public DNA databases containing germline antibody gene sequences. These germline sequences differ from mature antibody gene sequences by not fully including the assembled variable genes formed by V(D)J junctions during B cell maturation. Germline gene sequences also differ from those of high-affinity secondary antibodies in variable genes, typically centered in the CDRs. For example, somatic mutations are relatively rare in the amino-terminal portion of framework region 1 and the carboxy-terminal portion of framework region 4. Thus, it is not necessary to obtain the complete DNA sequence of a particular antibody to regenerate a complete recombinant antibody with a binding chemistry similar to that of the original antibody (see WO 99/45962). The heavy and light chain sequences partially spanning the CDR regions were used to determine germline variable and junctional gene segments involved in recombined antibody variable genes. The missing portions of the variable regions were then filled in with germline sequences. The heavy and light chain leader sequences are cleaved upon protein maturation and do not participate in the properties of the final antibody. To add missing sequences, cloned cDNA sequences were ligated with synthetic oligonucleotides by ligation or PCR amplification. Alternatively, complete variable regions can be synthesized as a set of short overlapping oligonucleotides and combined by PCR to generate complete synthetic variable region clones. This process has specific advantages, such as deletion or insertion of specific restriction sites, or optimization of specific codons.

来自杂交瘤的重链和轻链转录子的核苷酸序列可用于设计一组重叠的合成寡核苷酸而产生具有与天然序列相同的氨基酸编码能力的合成的V序列。合成的重链和κ链序列可在三个方面不同于天然序列：打 断重复核苷酸碱基的排列，以利于寡核苷酸合成和PCR扩增；根据Kozak的原则(Kozak，J.Biol.Chem.266，19867-19870(1991)引入优化的翻译起始位点；和在翻译起始位点上游构建HindIII位点。The nucleotide sequences of the heavy and light chain transcripts from the hybridomas can be used to design an overlapping set of synthetic oligonucleotides to generate a synthetic V sequence with the same amino acid encoding capacity as the native sequence. Synthetic heavy and kappa chain sequences can differ from natural sequences in three respects: the arrangement of repeated nucleotide bases is interrupted to facilitate oligonucleotide synthesis and PCR amplification; according to Kozak's principles (Kozak, J. Biol. Chem. 266, 19867-19870 (1991) introduction of an optimized translation initiation site; and construction of a HindIII site upstream of the translation initiation site.

对重链和轻链可变区而言，优化的编码和对应的非编码链序列在相应的非编码核苷酸寡核苷酸的中部打断为30-50个核苷酸。因此，对每条链而言，寡核苷酸可组装为重叠的双链形式，跨越150-400个核苷酸。然后以其作为模板，产生150-400个核苷酸的PCR扩增产物。典型地，单个可变区寡核苷酸组将打断为2组，分别扩增产生两个重叠的PCR产物。然后将这些重叠的产物通过PCR扩增连接，形成完整的可变区。在PCR扩增中也需要重链或轻链恒定区(包括κ轻链的BbsI位点，或γ重链的AgeI位点)的重叠片段，以产生可易于克隆到表达载体构建体中的片段。For the heavy and light chain variable regions, the optimized coding and corresponding noncoding strand sequences were interrupted by 30-50 nucleotides in the middle of the corresponding noncoding nucleotide oligonucleotides. Thus, oligonucleotides can assemble in overlapping double-stranded form, spanning 150-400 nucleotides per strand. Then use it as a template to generate a PCR amplification product of 150-400 nucleotides. Typically, a single variable region oligonucleotide set will be broken into 2 sets, amplified separately to generate two overlapping PCR products. These overlapping products are then joined by PCR amplification to form the complete variable region. Overlapping fragments of the heavy or light chain constant regions (including the BbsI site for the kappa light chain, or the AgeI site for the gamma heavy chain) are also required in PCR amplification to generate fragments that can be easily cloned into expression vector constructs .

然后将重新构建的重链和轻链可变区与克隆的启动子、引导序列、翻译起始区、恒定区3′非翻译区、多聚腺苷化和转录终止区、形成表达载体构建体的序列连接。重链和轻链表达构建体也可与单个载体共转染、顺序转染或单独转染到宿主细胞中，然后融合而从宿主细胞中表两条链。The reconstituted heavy and light chain variable regions were then combined with the cloned promoter, leader sequence, translation initiation region, constant region 3' untranslated region, polyadenylation and transcription termination region, to form an expression vector construct sequence connection. The heavy and light chain expression constructs can also be co-transfected with a single vector, sequentially transfected, or separately transfected into a host cell and then fused to express both chains from the host cell.

相似的流程可用于将新型抗原特异性移植到存在的成熟抗体中。典型地，受体抗体选自来源于相同的可变胚系基因作为CDR供体的抗体，但也可选择其它受体抗体。然后将供体抗体中的一个或单个CDRs通过上述技术进行转移。A similar procedure can be used to graft novel antigen specificity into existing mature antibodies. Typically, recipient antibodies are selected from antibodies derived from the same variable germline genes as the CDR donors, but other recipient antibodies may also be selected. One or individual CDRs in the donor antibody are then transferred by the techniques described above.

在本发明的一个技术方案中，-003和005和-024的结构特征用于产生结构相关的至少保留-003和005和-024的一个功能特性，即与CD3 8结合的抗CD38抗体，例如人抗CD3 8抗体。更具体地，可将-003和005和-024的一个或多个CDR区与已知的人框架区和CDRs连接，从而产生本发明的其它重组构建的人抗CD38抗体。In a technical solution of the present invention, the structural features of -003 and 005 and -024 are used to generate structurally related anti-CD38 antibodies that retain at least one functional property of -003, 005 and -024, that is, binding to CD38, for example Human anti-CD3 8 antibody. More specifically, one or more CDR regions of -003 and 005 and -024 can be linked to known human framework regions and CDRs to generate other recombinantly constructed human anti-CD38 antibodies of the invention.

用于构建人IgGκ的表达载体的示例质粒如下所述。构建质粒，使PCR扩增的Vκ重链和Vκ轻链cDNA序列用于重新构建完整的重链和轻链微基因。这些质粒可用于表达完整的人IgG1，κ或IgG4，κ抗体，类似的质粒可构建以表达其它重链同型体，或表达含λ轻链的抗体。Exemplary plasmids for constructing expression vectors for human IgGκ are as follows. Plasmids were constructed so that the PCR-amplified Vκ heavy chain and Vκ light chain cDNA sequences were used to reconstruct complete heavy and light chain minigenes. These plasmids can be used to express fully human IgG1, kappa or IgG4, kappa antibodies, and similar plasmids can be constructed to express other heavy chain isotypes, or to express antibodies containing lambda light chains.

本发明的CD38BPs，例如本发明的人抗CD38抗体可通过多种不同方式分离和鉴定，例如，可将筛选的杂交瘤在合适的烧瓶中培养以进行 单克隆抗体纯化。然后在用蛋白A-葡聚糖的亲和层析(对IgG1同型体抗体而言)(Pharmacia，Piscataway，NJ)或抗人IgG包被的葡聚糖，或对IgG3同型体抗体而言用蛋白G-葡聚糖进行亲和层析之前过滤上清，并进行浓缩。洗脱的IgG可通过凝胶电泳和高效液相色谱来检测以保证纯度。缓冲液可使用PBS，浓度可通过用1.43消光系数来测定OD₂₈₀。等分单克隆抗体并在-80℃保存。The CD38BPs of the present invention, such as the human anti-CD38 antibodies of the present invention, can be isolated and identified in various ways, for example, the screened hybridomas can be cultured in a suitable flask for monoclonal antibody purification. This was followed by affinity chromatography with protein A-dextran (for IgG1 isotype antibodies) (Pharmacia, Piscataway, NJ) or anti-human IgG-coated dextran, or for IgG3 isotype antibodies with The supernatant was filtered and concentrated before protein G-glucan affinity chromatography. Eluted IgG can be checked by gel electrophoresis and high performance liquid chromatography to ensure purity. PBS can be used as the buffer, and the concentration can be determined by using an extinction coefficient of 1.43 to determine the OD ₂₈₀ . Aliquot the monoclonal antibody and store at -80°C.

为了测定是否筛选的CD38BPs，例如人抗CD38单克隆抗体与特定的抗原决定簇结合，可使用定点或多点突变技术。To determine whether selected CD38BPs, such as human anti-CD38 monoclonal antibodies, bind to specific epitopes, site-directed or multiple point mutagenesis techniques can be used.

为了测定纯化抗体的同种型，可进行同种型ELISAs。微孔板的孔可用10μg/ml抗人Ig在4℃包被过夜。板用5％BSA(牛血清蛋白)封闭后，与10μg/ml单克隆抗体或纯化的同种型对照在室温下反应2个小时。然后将孔与人特异IgG1、IgG2、IgG3或IgG4、IgE、IgA1、IgA2或人IgM的偶联碱性磷酸酶的探针反应。洗涤后，用pNPP底物(1mg/ml)对板显色，并通过405nm处的OD进行分析。To determine the isotype of purified antibodies, isotype ELISAs can be performed. Wells of microplates can be coated with 10 μg/ml anti-human Ig overnight at 4°C. Plates were blocked with 5% BSA (bovine serum albumin) and reacted with 10 μg/ml monoclonal antibody or purified isotype control for 2 hours at room temperature. The wells were then reacted with alkaline phosphatase-conjugated probes specific for human IgG1, IgG2, IgG3 or IgG4, IgE, IgAl, IgA2 or human IgM. After washing, the plates were developed with pNPP substrate (1 mg/ml) and analyzed by OD at 405 nm.

可使用流式细胞仪来检测抗CD38抗体在免疫鼠的血清中的存在或CD38BPs(包括抗CD38抗体)与表达CD38的活细胞的结合。简言之，表达CD38的细胞系(在标准生长条件下生长)与各种浓度的溶在含0.1％BSA和0.02％叠氮化钠的PBS中的CD38BP混合，在4℃温育30分钟。洗涤后，将细胞与荧光素标记的抗人IgG抗体在与一抗染色相同的条件下进行反应。可用流式细胞仪(例如Becton Dickinson FACS仪)通过流式细胞仪对样品进行分析，以日光和侧向光散射特性来对单个活细胞设门。可使用通过荧光显微镜技术的方法(除外或替代)流式细胞仪方法。可如上所述对细胞进行染色，并通过荧光显微镜进行检测。该方法可观察单个细胞，但根据抗原密度可能降低灵敏度。Flow cytometry can be used to detect the presence of anti-CD38 antibodies in the sera of immunized mice or the binding of CD38BPs (including anti-CD38 antibodies) to live cells expressing CD38. Briefly, CD38-expressing cell lines (grown under standard growth conditions) were mixed with various concentrations of CD38BP in PBS containing 0.1% BSA and 0.02% sodium azide and incubated at 4°C for 30 minutes. After washing, cells were reacted with fluorescein-labeled anti-human IgG antibody under the same conditions as for primary antibody staining. Samples can be analyzed by flow cytometry using a flow cytometer (eg, a Becton Dickinson FACS instrument), gating on individual living cells on sunlight and side light scatter properties. Methods by fluorescence microscopy (in addition to or instead of) flow cytometric methods may be used. Cells can be stained as described above and examined by fluorescence microscopy. This method allows visualization of single cells, but sensitivity may be reduced depending on antigen density.

CD38BPs，例如CD38人IgGs可进一步通过Western印迹检测对CD38抗原的反应性。简言之，制备表达CD38细胞的细胞提取物，并进行十二烷基磺酸钠(SDS)聚丙烯酰胺凝胶电泳。电泳后，将分离的抗原转移到硝酸纤维素膜上，用20％脱脂奶粉封闭，并用待测的CD38BPs进行检测。可用抗人IgG碱性磷酸酶来检测人IgG的结合，用BCIP/NBT底物片(SigmaChem.Co.，St.Louis，MO)进行显色，但也可用CD38BP其它特异部分来直接检测。CD38BPs, such as CD38 human IgGs, can be further tested for reactivity to CD38 antigen by Western blotting. Briefly, cell extracts of CD38-expressing cells were prepared and subjected to sodium dodecylsulfate (SDS) polyacrylamide gel electrophoresis. After electrophoresis, the separated antigens were transferred to nitrocellulose membranes, blocked with 20% nonfat dry milk, and detected with the CD38BPs to be tested. Binding of human IgG can be detected with anti-human IgG alkaline phosphatase, developed with BCIP/NBT substrate sheets (SigmaChem. Co., St. Louis, MO), but can also be directly detected with other specific portions of CD38BP.

除了与CD38特异结合外，可检测CD38BPs(包括人抗CD38抗体) 对表达CD38的细胞的各种活性，例如，但不限定于胰岛素产生、Ca²⁺ 释放、细胞因子产生、裂解诱导作用、分化和增殖的抑制作用。In addition to specific binding to CD38, various activities of CD38BPs (including human anti-CD38 antibodies) on CD38-expressing cells can be detected, such as, but not limited to, insulin production, Ca ²⁺ release, cytokine production, lysis induction, differentiation and inhibition of proliferation.

在一个技术方案中，本发明提供可表达特异结合CD38的人抗体的转基因和转染色体非人动物，例如转基因或转染色体鼠。在一个特定技术方案中，本发明提供具有含人重链转基因的基因组的转基因或转染色体鼠，当用表达CD38的细胞免疫时，该鼠可产生人抗CD38抗体。在转基因情况下，人重链转基因可整合到鼠染色体DNA中，例如HuMAb鼠，如下详细描述。选择性地，在转染色体鼠(例如KM)情况下，人重链转基因可在染色体外维持，如WO02/43478所述。这种转基因和转染色体动物可通过V-D-J/V-J重组和同种型转换而产生多种针对CD38的人单克隆抗体的同种型(例如IgG、IgA和/或IgE)。对具有异源抗体库的外源抗原刺激应答的转基因或转染色体非人动物的设计需要转基因动物中所含的异源免疫球蛋白转基因在B细胞发育过程中正确行使功能。这包括，例如异源重链转基因的同种型转换。因此，构建转基因，使其可诱导同种型转换及具有一个或多个以下抗体基因的特征：(1)高水平和细胞类型特异的表达，(2)功能性基因重排，(3)激活并对等位基因排除有应答，(4)表达足够的一级库，(5)信号转导，(6)体细胞高突变，及(7)在免疫应答中转基因抗体座位是显性的。In one technical solution, the present invention provides transgenic and transchromosomal non-human animals expressing human antibodies that specifically bind to CD38, such as transgenic or transchromosomal mice. In a specific embodiment, the invention provides a transgenic or transchromosomal mouse having a genome comprising a human heavy chain transgene, which mouse produces human anti-CD38 antibodies when immunized with cells expressing CD38. In the transgenic case, the human heavy chain transgene can be integrated into the chromosomal DNA of a mouse, eg a HuMAb mouse, as described in detail below. Alternatively, in the case of transchromosomal mice (eg KM), the human heavy chain transgene can be maintained extrachromosomally, as described in WO02/43478. Such transgenic and transchromosomal animals can produce various isotypes (eg, IgG, IgA and/or IgE) of human monoclonal antibodies directed against CD38 by V-D-J/V-J recombination and isotype switching. The design of transgenic or transchromosomal non-human animals that respond to foreign antigen stimulation with a heterologous antibody repertoire requires that the heterologous immunoglobulin transgenes contained in the transgenic animals function correctly during B cell development. This includes, for example, isotype switching of heterologous heavy chain transgenes. Therefore, transgenes were constructed to induce isotype switching and to be characterized by one or more of the following antibody genes: (1) high-level and cell-type-specific expression, (2) functional gene rearrangement, (3) activation and respond to allelic exclusion, (4) express sufficient primary repertoire, (5) signal transduction, (6) somatic hypermutation, and (7) are dominant in the transgenic antibody locus in the immune response.

并不是所有前述标准都满足。例如，在某些技术方案中，其中转基因动物的内源免疫球蛋白座位是被功能上破坏的，转基因不需激活等位基因排除反应。另外，在某些技术方案中，其中转基因含功能性重组的重链和/或轻链免疫球蛋白基因，因此功能性基因重组的第二个标准是不需要的，至少对转基因已经重组的情况而言。对于分子免疫学背景，参见，Fundamental Immunology，2nd edition(1989)、Paul William E.，ed.Raven Press，N.Y。Not all of the foregoing criteria are met. For example, in certain embodiments in which the endogenous immunoglobulin loci of the transgenic animal are functionally disrupted, the transgene need not activate an allelic exclusion response. In addition, in certain technical solutions, where the transgene contains functionally recombined heavy and/or light chain immunoglobulin genes, the second criterion of functional genetic recombination is not required, at least for cases where the transgene has been recombined In terms of. For background in molecular immunology see, Fundamental Immunology, 2nd edition (1989), Paul William E., ed. Raven Press, N.Y.

在特定技术方案中，转基因或转染色体非人动物用于在转基因动物的胚系中产生含重组的、未重组的或同时含有重组和未重组的异源免疫球蛋白重链和轻链转基因的本发明的人单克隆抗体。每种重链转基因含有至少一个C_H基因。另外，重链转基因可含有功能性同型体转换序列，可支持在转基因动物的B细胞中编码多个C_H基因的异源转基因的同种型转换。这种转换序列是那些在作为转基因C_H基因来源的物种中的胚系免疫球蛋白座位中天然存在的序列，或这种转换序列来源于那些在接 受转基因构建体的物种(转基因动物)中存在的序列。例如，用于产生转基因鼠的人转基因构建体如果整合了那些与鼠重链座位天然存在的序列相似的转换序列，则可产生高频率的同种型转换事件，可能鼠转换序列优先对鼠转换重组酶系统起作用，而人转换序列不是这样。可通过常规克隆方法分离和克隆转换序列，或从基于免疫球蛋白转换区序列相关的公布的序列信息设计的重叠的合成寡核苷酸从头合成(Mills et al.，Nucl.Acids Res.15，7305-7316(1991)Sideras et al.，Intl.Immunol.1，631-642(1989))。对每种前述转基因动物而言，在转基因动物的B细胞中发现大部分有功能的重组异源重链和轻链免疫球蛋白转基因(至少10％)。In a specific technical solution, a transgenic or transchromosomal non-human animal is used to produce recombinant, unrecombined, or both recombinant and unrecombined heterologous immunoglobulin heavy and light chain transgenes in the germline of the transgenic animal. Human monoclonal antibodies of the invention. Each heavy chain transgene contains at least one _CH gene. In addition, the heavy chain transgene can contain a functional isotype switching sequence that can support isotype switching of heterologous transgenes encoding multiple _CH genes in the B cells of the transgenic animal. Such switch sequences are those found naturally at germline immunoglobulin loci in the species from which the transgenic _CH gene was derived, or such switch sequences are derived from those present in the species receiving the transgenic construct (transgenic animal) the sequence of. For example, human transgene constructs used to generate transgenic mice that incorporate switch sequences that are similar to those naturally occurring at the murine heavy chain locus can generate high frequencies of isotype switching events, presumably the murine switch sequences preferentially switch to the murine The recombinase system works whereas the human switch sequence does not. The switch sequence can be isolated and cloned by conventional cloning methods, or synthesized de novo from overlapping synthetic oligonucleotides designed based on published sequence information related to immunoglobulin switch region sequences (Mills et al., Nucl. Acids Res. 15, 7305-7316 (1991) Sideras et al., Intl. Immunol. 1, 631-642 (1989)). For each of the aforementioned transgenic animals, the majority of functional recombinant heterologous heavy and light chain immunoglobulin transgenes (at least 10%) were found in the B cells of the transgenic animals.

用于产生本发明的转基因非人动物的转基因包括含编码至少一个可变基因片段、一个多样性基因片段、一个连接基因片段和至少一个恒定区基因片段的DNA的重链转基因。免疫球蛋白轻链转基因含有编码至少一个可变基因片段、一个连接基因片段和至少一个恒定区基因片段的DNA。编码轻链和重链基因片段的基因片段对转基因动物是异源的，因为它们来源于或对应于编码来自不组成转基因非人动物的物种的免疫球蛋白重链和轻链基因片段的DNA。在本发明的一个技术方案中，构建转基因，使单个基因片段不重组，即不重排以编码功能性免疫球蛋白轻链或重链。这种未重排的转基因可支持V、D和J基因片段的重组(功能性重组)，而且支持当暴露到CD38抗原中，所有或部分D区整合在转基因动物中所得到的重组免疫球蛋白重链中。The transgenes used to generate the transgenic non-human animals of the invention include heavy chain transgenes comprising DNA encoding at least one variable gene segment, one diversity gene segment, one junction gene segment, and at least one constant region gene segment. The immunoglobulin light chain transgene contains DNA encoding at least one variable gene segment, one junction gene segment and at least one constant region gene segment. The gene segments encoding the light and heavy chain gene segments are heterologous to the transgenic animal in that they are derived from or correspond to DNA encoding the immunoglobulin heavy and light chain gene segments from the species that does not make up the transgenic non-human animal. In one technical solution of the present invention, the transgene is constructed so that a single gene segment does not recombine, that is, does not rearrange to encode a functional immunoglobulin light chain or heavy chain. This unrearranged transgene supports recombination of the V, D, and J gene segments (functional recombination) and supports the resulting recombinant immunoglobulin in the transgenic animal when all or part of the D region is integrated when exposed to the CD38 antigen in the heavy chain.

在选择的技术方案中，转基因含有未重组的“微座位”。这种转基因典型地含有C、D和J片段的实质部分，及V基因片段的亚组。在这种转基因构建体中，各种调控序列，例如启动子、增强子、种类转换区、RNA加工中的拼接供体和拼接受体序列、重组信号等均含有来自异源DNA的相应序列。这种调控序列可整合到来自本发明所用的非人动物相同或相关的物种中的转基因中。例如，人免疫球蛋白基因片段可与转基因中用于转基因鼠的啮齿类免疫球蛋白增强子序列连接。选择性地，合成的调控序列可整合在转基因中，其中这种合成的调控序列不与已知天然存在于哺乳动物基因组中的功能DNA序列同源。合成的调控序列根据同一性原则进行设计，例如，那些特异针对拼接受体位点或启动子/增强子元件的允许序列。例如，与天然存在的胚系Ig座位相比较，微座 位含有至少一个内部(即不在部分的末端)缺失非必需DNA部分(例如，干扰序列；内含子或其内含子或部分)的基因组免疫球蛋白座位。In a selected technical solution, the transgene contains unrecombined "microlocus". Such transgenes typically contain substantial portions of the C, D and J segments, and a subset of the V gene segments. In such transgenic constructs, various regulatory sequences, such as promoters, enhancers, species switching regions, splice donor and splice acceptor sequences in RNA processing, recombination signals, etc., all contain corresponding sequences from heterologous DNA. Such regulatory sequences may be incorporated into a transgene from the same or a related species of the non-human animal used in the invention. For example, a human immunoglobulin gene segment can be linked to a rodent immunoglobulin enhancer sequence in a transgene for a transgenic mouse. Alternatively, synthetic regulatory sequences may be incorporated into the transgene, where such synthetic regulatory sequences are not homologous to functional DNA sequences known to occur naturally in mammalian genomes. Synthetic regulatory sequences are designed on the basis of identity principles, for example, permissive sequences that are specific for splicing acceptor sites or promoter/enhancer elements. For example, a microlocus contains at least one genome in which nonessential DNA portions (e.g., interfering sequences; introns or introns or portions thereof) are deleted internally (i.e., not at the end of the portion) compared to naturally occurring germline Ig loci Immunoglobulin loci.

转基因和转染色体非人动物例如鼠的实施例具有显著的免疫球蛋白产生的库，理想地，在调整体积后与人的库实质上相似。Examples of transgenic and transchromosomal non-human animals such as murines have remarkably immunoglobulin-producing repertoires, ideally substantially similar to human repertoires after adjusting for volume.

在调整体积后，人的库理想地通常密度至少约为10％或更高，例如25％到50％或更多。通常，构建引入到鼠基因组中的V、J和D区的不同数目及V(-D-)J基因片段重组的其它多样性和连接区的随机核苷酸增加，可产生至少约1000个不同的免疫球蛋白(理想地为IgG1)，例如10⁴到10⁶或更多。典型地，免疫球蛋白对预选的抗原的亲和性低于10^-8M，例如低于10^-9M、10^-10 M或10^-11M，或甚至更低。如上所述的转基因和转染色体非人动物，例如鼠可用例如表达Cd38的细胞免疫。选择性地，转基因动物可用编码人CD38的DNA免疫。然后动物产生B细胞，并通过转换重组(顺式转换)进行种类转换，并表达与CD38反应的免疫球蛋白。免疫球蛋白是人抗体(也称为“人序列抗体”)，其中重链和轻链多肽由包括体细胞突变衍生的序列和V区重组连接序列及胚系编码序列的人转基因序列编码；这些人抗体是指实质上与人V_L 和JL或V_H、D_H和JH基因片段编码的多肽序列相同，即使由于体细胞突变和不同的V-J和V-D-J重组连接而存在其它非胚系序列。每个抗体链的可变区典型地与胚系V和J基因片段同源性为80％，对重链而言，人胚系V、D和J基因片段通常与转基因中存在的人胚系序列的同源性至少为85％；通常90或95％或更高。但由于非胚系序列通过体细胞突变和VJ和VDJ连接而被引入，因此人序列抗体通常具有某些不被在鼠的胚系中的人转基因中的人V、D或J基因片段所编码的某些可变区序列。典型地，这种非胚系序列(或单个核苷酸位置)将成簇存在并邻近CDRs，或在簇中的体细胞突变是已知的区域。After volume adjustment, the human pool desirably typically has a density of at least about 10% or higher, such as 25% to 50% or more. Typically, different numbers of V, J, and D regions introduced into the murine genome by construction and other diversity in recombination of V(-D-)J gene segments and random nucleotide additions in junctional regions can produce at least about 1000 different of immunoglobulin (ideally IgG1), such as 10 ⁴ to 10 ⁶ or more. Typically, the immunoglobulin has an affinity for the preselected antigen of less than 10 ⁻⁸ M, such as less than 10 ⁻⁹ M, 10 ⁻¹⁰ M or 10 ⁻¹¹ M, or even lower. Transgenic and transchromosomal non-human animals such as mice, as described above, can be immunized, for example, with cells expressing Cd38. Alternatively, transgenic animals can be immunized with DNA encoding human CD38. The animal then generates B cells that undergo class switching through switch recombination (cis-switching) and express immunoglobulins reactive with CD38. Immunoglobulins are human antibodies (also referred to as "human sequence antibodies") in which the heavy and light chain polypeptides are encoded by human transgenic sequences including sequences derived from somatic mutations and recombined junctional V region and germline coding sequences; these Human antibodies refer to polypeptide sequences substantially identical to human _VL and JL or _VH , _DH , and JH gene segments encoded by human antibodies, even if other non-germline sequences are present due to somatic mutations and different VJ and VDJ recombination linkages. The variable regions of each antibody chain are typically 80% homologous to germline V and J gene segments, and for the heavy chains, human germline V, D, and J gene segments are usually identical to the human germline gene segments present in the transgene. The sequence identity is at least 85%; usually 90 or 95% or more. However, because non-germline sequences are introduced by somatic mutation and VJ and VDJ joining, human sequence antibodies usually have certain human V, D, or J gene segments not encoded by human transgenes in the germline of the mouse certain variable region sequences. Typically, such non-germline sequences (or single nucleotide positions) will be present in clusters adjacent to CDRs, or in regions where somatic mutations in clusters are known.

本发明也提供来源于此处所述的转基因或转染色体非人动物的B细胞。B细胞可用于产生表达与人CD38以高亲和性(例如，解离平衡常数(K_D)低于10^-8M)结合的人单克隆抗体的杂交瘤。因此，在一个技术方案中，本发明提供产生亲和性(K_D)低于10^-8M，例如低于10^-9M、10^-10M或10^-11M，或甚至更低的人抗体，用放射性标记的单克隆抗体通过斯卡查德分析或通过FACS分析测定最大半衰期结合浓度、或通过表面等离子共振分析在BIAcore仪器上进行测定。本发明提供含由 以下组成的抗CD38抗体，其中人序列轻链含有(1)具有实质上与由人V_L基因片段和人J_L片段编码的多肽序列相同的多肽序列的轻链可变区，及(2)由人C_L基因片段编码的轻链恒定区；人序列重链含有(1)具有实质上与由人V_H基因片段、D区和人J_H片段编码的多肽序列相同的多肽序列的轻链可变区，及(2)由人CH基因片段编码的轻链恒定区。应注意人D基因实质上通过重组和体细胞突变事件而发生改变，从而可能不易于识别原来的人胚系序列。The invention also provides B cells derived from the transgenic or transchromosomal non-human animals described herein. B cells can be used to generate hybridomas expressing human monoclonal antibodies that bind to human CD38 with high affinity (eg, dissociation equilibrium constant ( _KD ) below 10 ⁻⁸ M). Thus, in one embodiment, the present invention provides for the generation of human compounds having an affinity (K _D ) below 10 ⁻⁸ M, such as below 10 ⁻⁹ M, 10 ⁻¹⁰ M or 10 ⁻¹¹ M, or even lower. Antibodies, maximal half-life binding concentrations were determined by Scatchard analysis or by FACS analysis with radiolabeled monoclonal antibodies, or by surface plasmon resonance analysis on a BIAcore instrument. The present invention provides an anti-CD38 antibody comprising a human sequence light chain comprising (1) a light chain variable region having a polypeptide sequence substantially identical to a polypeptide sequence encoded by a human _VL gene segment and a human _JL segment , and (2) the light chain constant region encoded by the human _CL gene segment; the human sequence heavy chain contains (1) a polypeptide sequence substantially identical to the polypeptide sequence encoded by the human _VH gene segment, the D region and the human _JH segment The light chain variable region of the polypeptide sequence, and (2) the light chain constant region encoded by the human CH gene fragment. It should be noted that the human D gene is substantially altered by recombination and somatic mutation events, so that the original human germline sequence may not be readily recognizable.

针对CD38的高亲和性人单克隆抗体的开发可通过扩展具有含整合的人免疫球蛋白转基因的基因组的转基因非人动物中的人可变区基因片段的文库的方法来进行，所述方法包括，将含不存在于所述整合的人免疫球蛋白转基因中的V区基因片段的V基因转基因整合到基因组中。通常，V区转基因是含有部分在人基因组中天然存在或通过重组方法拼接在一起的人V_H或V_L(Vκ)基因片段阵列的酵母人工染色体(YAC)，这些片段包括无序的或省略的V基因片段。通常至少在YAC中含5个或更多功能性V基因片段。在这种变化中，可通过V文库扩展方法制备转基因动物，其中动物表达含有由存在于V区转基因上的V区基因片段编码的可变区序列和人Ig转基因上编码的C区的免疫球蛋白链。通过V文库扩展方法，可产生具有至少5个不同V基因的转基因动物；例如含至少约34个V基因或更多的动物。某些V基因片段可能是没有功能的(例如，假基因等)；如果需要，这些片段可通过技术人员已知的重组方法被保留或选择性删除。Development of high-affinity human monoclonal antibodies directed against CD38 can be performed by a method of expanding libraries of human variable region gene fragments in transgenic non-human animals having genomes containing integrated human immunoglobulin transgenes Including, integrating into the genome a V gene transgene containing a V region gene segment not present in the integrated human immunoglobulin transgene. Typically, the V region transgene is a yeast artificial chromosome (YAC) containing an array of human _VH or _VL (Vκ) gene segments, including disordered or omitted The V gene fragment. Usually at least five or more functional V gene segments are contained in the YAC. In this variation, transgenic animals can be prepared by the V library expansion method, wherein the animal expresses an immunoglobulin containing the variable region sequences encoded by the V region gene segment present on the V region transgene and the C region encoded on the human Ig transgene protein chain. Through the V library expansion method, transgenic animals can be generated with at least 5 different V genes; for example, animals containing at least about 34 V genes or more. Certain V gene segments may be non-functional (eg, pseudogenes, etc.); if desired, these segments can be retained or selectively deleted by recombination methods known to the skilled artisan.

当构建含具有扩展的V片段文库的功能性YAC的鼠胚系，实质上不存在含J和C基因片段的人Ig转基因时，则可扩增该特性，并扩增到其它遗传背景，包括其中具有扩展的V片段文库的功能性YAC导入到具有不同的人Ig转基因的非人动物胚系的背景。多个具有扩展的V片段文库的功能性YACs可导入到胚系中，使其与人Ig转基因(或多个人Ig转基因)一起起作用。虽然此处是指YAC转基因，但如果这种转基因整合到实质上缺失酵母序列的基因组中时，例如在酵母中需自主复制序列中；则这种序列在酵母复制后不再需要时(即，在导入到鼠ES细胞或鼠原接合子(prozygote)中之前)，可选择性地通过基因工程(例如，限制性消化和脉冲凝胶电泳或其它合适的方法)除去。扩增人序列免疫球蛋白表达的方法包括培养具有人Ig转基因，选择性地也具有含 扩展的V片段文库的功能性YAC的转基因动物。V_H和V_L基因片段均可存在于YAC中。转基因动物可在任何实验人员所需的背景中培养，包括含有其它人转基因，包括人Ig转基因和/或编码其它人淋巴细胞蛋白的转基因的背景。本发明也提供由具有扩展的V区文库YAC转基因产生的高亲和性人序列免疫球蛋白。虽然以上描述了本发明的转基因动物的特定技术方案，其它划分在以下3类中的技术方案是期望的：This property can be amplified when a murine germline containing a functional YAC with an expanded V segment library is constructed in the substantial absence of human Ig transgenes containing J and C gene segments, and extended to other genetic backgrounds, including Background in which a functional YAC with an expanded V-segment library was introduced into the germline of a non-human animal with a different human Ig transgene. Multiple functional YACs with an expanded V-segment library can be introduced into the germline to function with a human Ig transgene (or multiple human Ig transgenes). Although a YAC transgene is referred to here, if such a transgene is integrated into a genome that is substantially devoid of yeast sequences, such as in yeast where autonomously replicating sequences are required; then such sequences are no longer required after yeast replication (i.e., prior to introduction into murine ES cells or murine prozygotes) can optionally be removed by genetic engineering (eg, restriction digestion and pulsed gel electrophoresis or other suitable methods). Methods of amplifying expression of human sequence immunoglobulins include culturing transgenic animals with human Ig transgenes, optionally also functional YACs containing expanded V-segment libraries. Both _VH and _VL gene segments can be present in a YAC. Transgenic animals can be raised in any background desired by the experimenter, including backgrounds containing other human transgenes, including human Ig transgenes and/or transgenes encoding other human lymphocyte proteins. The invention also provides high affinity human sequence immunoglobulins produced from YAC transgenes with expanded V region libraries. Although the specific technical solutions of the transgenic animals of the present invention have been described above, other technical solutions classified in the following 3 categories are desirable:

I.含未重组重链和重组轻链免疫球蛋白转基因的转基因动物；I. Transgenic animals containing non-recombined heavy chain and recombinant light chain immunoglobulin transgenes;

II.含未重组重链和未重组轻链免疫球蛋白转基因的转基因动物；及II. Transgenic animals containing unrecombined heavy chain and unrecombined light chain immunoglobulin transgenes; and

III.含重组重链和未重组轻链免疫球蛋白转基因的转基因动物。III. Transgenic animals containing recombinant heavy chain and unrecombined light chain immunoglobulin transgenes.

在一个技术方案中，本发明提供含治疗有效量的本发明的CD38BP的药物组合物。药物组合物可由药学上可接受的载体或稀释剂和其它已知的佐剂和赋形剂通过例如在Remington：The Science and Practice ofPharmacy，19th Edition，Gennaro，Ed.，MackPublishing Co.，Easton，PA1 1995中描述的传统技术制备。In one technical solution, the present invention provides a pharmaceutical composition containing a therapeutically effective amount of the CD38BP of the present invention. Pharmaceutical compositions can be formulated with pharmaceutically acceptable carriers or diluents and other known adjuvants and excipients as described, for example, in Remington: The Science and Practice of Pharmacy, 19th Edition, Gennaro, Ed., MackPublishing Co., Easton, PA 1 Prepared by conventional techniques described in 1995.

药学上可接受的载体或稀释剂和任何其它已知的佐剂和赋形剂对本发明所选的化合物和所选的给药方式是合适的。载体和其它药物组合物的成分的合适性是根据对本发明所选的化合物或药物组合物所需的生物学特性在抗原结合上没有显著的负面影响而确定的(例如，小于实质影响(小于10％或更低的相对抑制作用，5％或更低的相对抑制作用等)。A pharmaceutically acceptable carrier or diluent and any other known adjuvants and vehicles are suitable for the selected compound of the invention and the selected mode of administration. The suitability of the carrier and other ingredients of the pharmaceutical composition is based on the fact that there is no significant negative effect on antigen binding (e.g., less than a substantial effect (less than 10 % or less relative inhibition, 5% or less relative inhibition, etc.).

本发明的药物组合物也可包括稀释剂、填充剂、盐、缓冲液、去垢剂(例如非离子去垢剂，例如Tween80)、稳定剂、稳定剂(例如糖或无蛋白的氨基酸)、防腐剂、组织固定剂和/或其它适于包含在药物组合物中的材料。The pharmaceutical compositions of the present invention may also include diluents, fillers, salts, buffers, detergents (e.g. non-ionic detergents such as Tween 80), stabilizers, stabilizers (e.g. sugars or protein-free amino acids), Preservatives, tissue fixatives and/or other materials suitable for inclusion in pharmaceutical compositions.

可改变本发明的药物组合物中的活性成分的实际剂量水平，以获得对特定患者、组成和给药模式而言，可达到所需治疗性应答的有效的活性成分的量，但对患者是无害的。选择的剂量水平依赖于各种药物动力学因素，包括所用的本发明的药物组合物或其酯、盐或胺的特定组成的活性、给药途径、给药时间、所用特定化合物排泄速度、治疗持续时间、与所用特定组分联合使用的其它的药物、化合物和/或材料、年龄、性别、体重、疾病症状、被治疗患者的一般健康情况和药物史及医学领域已知 的其它因素。Actual dosage levels of the active ingredients in the pharmaceutical compositions of this invention may be varied to obtain an effective amount of the active ingredient to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, but the patient's harmless. The selected dosage level will depend on various pharmacokinetic factors, including the activity of the particular composition of the pharmaceutical composition of the present invention or its ester, salt or amine employed, the route of administration, the time of administration, the rate of excretion of the particular compound employed, the therapeutic Duration, other drugs, compounds and/or materials used in conjunction with the particular components used, age, sex, weight, disease symptoms, general health and drug history of the patient being treated and other factors known in the medical field.

药物组合物可通过任何合适的途径和方式给药。合适的给药本发明的化合物的体内和体外途径在本领域是已知的，本领域技术人员可进行选择。Pharmaceutical compositions can be administered by any suitable route and manner. Suitable in vivo and in vitro routes of administration of the compounds of the invention are known in the art and can be selected by one skilled in the art.

本发明的化合物可通过任何合适的途径，例如口服、鼻服、吸入式、局部(包括口腔、经皮肤和舌下)、直肠、阴道和/或肠胃外途径。The compounds of the invention may be administered by any suitable route, eg, oral, nasal, inhaled, topical (including buccal, transdermal and sublingual), rectal, vaginal and/or parenteral.

在一个技术方案中，本发明的药物组合物通过例如与惰性稀释剂或同化的可食用载体进行口服给药。活性成分可包括在硬的或软的壳明胶胶囊中、压缩为片剂、或直接混合在患者的饮食中。适于口服给药的本发明的药物组合物包括含本领域已知的合适载体的可吸收片剂、口腔片剂、药片、胶囊、甘香洒剂、悬浮液、糖浆、糯米纸等。为了对本发明的化合物进行口服给药，需要将化合物包被在可抑制其失活的化合物中或共同给药。In one technical solution, the pharmaceutical composition of the present invention is administered orally, eg, with an inert diluent or an assimilable edible carrier. The active ingredient can be included in hard or soft shell gelatin capsules, compressed into tablets, or mixed directly into the patient's diet. The pharmaceutical composition of the present invention suitable for oral administration includes ingestible tablets, buccal tablets, troches, capsules, sweeteners, suspensions, syrups, wafers and the like containing suitable carriers known in the art. In order to administer the compound of the present invention orally, it is necessary to coat the compound in a compound that inhibits its inactivation or co-administer it.

在一个技术方案中，本发明的药物组合物是通过鼻给药的。适于经鼻给药的本发明的药物组合物在本领域是已知的，典型地包括喷剂、鼻滴剂和吸入剂。In one embodiment, the pharmaceutical composition of the invention is administered nasally. Pharmaceutical compositions of the invention suitable for nasal administration are known in the art and typically include sprays, nasal drops and inhalants.

在一个技术方案中，本发明的药物组合物是局部给药的。适于局部或经皮肤给药的本发明的药物组合物包括含本领域已知的合适载体的粉剂、喷剂、药膏、泥膏、乳剂、洗剂、凝胶、溶液、片剂和吸入剂。In one embodiment, the pharmaceutical composition of the invention is administered topically. Pharmaceutical compositions of the present invention suitable for topical or transdermal administration include powders, sprays, salves, poultices, creams, lotions, gels, solutions, tablets and inhalants with suitable carriers known in the art. .

在一个技术方案中，本发明的药物组合物是直肠给药的。适于直肠给药的本发明的药物组合物在本领域是已知的，包括凝胶、泥膏、喷雾配方、栓剂。In one embodiment, the pharmaceutical composition of the invention is administered rectally. Pharmaceutical compositions of the present invention suitable for rectal administration are known in the art and include gels, poultices, spray formulations, suppositories.

在一个技术方案中，本发明的药物组合物是阴道给药的。适于阴道给药的本发明的药物组合物包括含本领域已知的合适载体的阴道栓剂、止血垫、乳剂、凝胶、泥膏、泡沫或喷雾配方。In one embodiment, the pharmaceutical composition of the present invention is administered vaginally. Pharmaceutical compositions of the present invention suitable for vaginal administration include pessaries, tampons, creams, gels, poultices, foams or spray formulations with suitable carriers known in the art.

在一个技术方案中，本发明的药物组合物是肠胃外给药的。In one embodiment, the pharmaceutical composition of the invention is administered parenterally.

此处所用术语“肠胃外给药”和“在肠胃外给药”是指除肠和局部给药外的给药方式，通常是通过注射进行，包括表皮、静脉内、肌肉内、动脉内、玻璃体内、眼窝内、心脏内、皮内、腹膜内、腱内、经气管、皮下、表皮下、关节内、囊下、蛛网膜下、脊柱内、头颅内、胸内、硬脑膜外和层间注射和灌液。The terms "parenteral administration" and "parenteral administration" as used herein refer to modes of administration other than enteral and topical administration, usually by injection, including epidermal, intravenous, intramuscular, intraarterial, Intravitreal, intraorbital, intracardiac, intradermal, intraperitoneal, intratendinous, transtracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, intracranial, intrathoracic, epidural and laminar Between injection and irrigation.

在一个技术方案中，药物组合物通过静脉内或皮下注射或灌液给 药。在一个技术方案中，本发明的化合物以晶体形式通过皮下注射给药，可与Yang et al.，PNAS USA 100(12)，6934-6939(2003)比较。In one technical solution, the pharmaceutical composition is administered by intravenous or subcutaneous injection or perfusion. In one embodiment, the compound of the present invention is administered in crystalline form by subcutaneous injection, compare with Yang et al., PNAS USA 100(12), 6934-6939 (2003).

药物组合物可用本领域已知的医疗设备给药。例如，在一个技术方案中，本发明的药物组合物可用无针的皮下组织注射设备，例如在US5,399,163、US 5,383,851、US 5,312,335、US 5,064,413、US 4,941,880、US 4,790,824或US 4,596,556中描述的设备给药。用于本发明的已知灌液和模式的实施例包括：US 4,487,603描述了灌液用微注射泵以可控速率用于药物分散；US 4,486,194描述了通过皮肤给药药物的治疗性设备；US 4,447,233描述了以精确的灌液速率递送药物的药物灌输泵；US4,447,224描述了进行持续药物递送的流式可植入灌液设备；US4,439,196描述了具有多室间隔区的渗透性药物递送系统，及US4,475,196描述了渗透性药物递送系统。许多其它这样的灌输、递送系统和模式对本领域技术人员是已知的。Pharmaceutical compositions can be administered using medical devices known in the art. For example, in one embodiment, the pharmaceutical composition of the present invention may be administered with a needle-free hypodermic tissue injection device such as those described in US 5,399,163, US 5,383,851, US 5,312,335, US 5,064,413, US 4,941,880, US 4,790,824 or US 4,596,556 medication. Examples of known irrigation solutions and modes for use in the present invention include: US 4,487,603 describes irrigation with a microsyringe pump for drug dispersion at a controlled rate; US 4,486,194 describes a therapeutic device for administering drugs through the skin; US 4,447,233 describes a drug infusion pump that delivers drug at a precise infusion rate; US 4,447,224 describes a flowable implantable infusion device for sustained drug delivery; US 4,439,196 describes osmotic drug delivery with multiple compartments system, and US 4,475,196 describes an osmotic drug delivery system. Many other such infusion, delivery systems and modes are known to those skilled in the art.

本发明的药物组合物可制备为用于特定的给药途径，例如口服、鼻服、局部(包括口腔、皮肤和舌下)、直肠、阴道和/或肠胃外给药。药物组合物可方便的以单位剂量形式存在，可通过任何药学领域已知的方法来制备。可与载体材料联合使用以产生单剂量形式的活性成分的量可根据被治疗患者和特定的给药方式而改变。可与载体材料联合使用以产生单剂量形式的活性成分的量通常是可产生治疗性效果的组分的量。通常，以100％表示，该量的范围从约0.01％到约99％的活性成分，例如从约0.1％到约70％，例如从约1％到约30％。不管所选的给药途径，可作为药学上可接受的盐或溶解性水合形式的本发明的化合物和/或本发明的药物组合物通过本领域技术人员已知的常规方法配制成药学上可接受的剂量形式。“药学上可接受的盐”是指可保持母化合物所需生物学活性而没有不需的毒性作用的盐(参见，例如Berge，S.M.et al.，J.Pharm.Sci.66，1-19(1977))。这种盐的实施例包括加酸盐和加碱盐。加酸盐包括那些来源于非毒性无机酸，例如盐酸、硝酸、类似、硫酸、氢溴酸、氢碘酸、磷酸等，以及来自无毒的有机酸，例如脂肪族单和二羧基酸、苯基取代的链烷酸、羟基链烷酸、芳香族酸、脂肪族和芳香族磺酸等的盐。碱加盐包括那些来源于碱土金属，例如钠、钾、镁、钙等，以及来源于无毒的有机胺，例如N.N′-二苄基乙二胺、N-甲基葡糖胺、氯普鲁卡因、维生素B复合物、二乙醇胺、乙(撑)二胺、普鲁卡因等 的盐。The pharmaceutical compositions of the present invention may be formulated for a particular route of administration, such as oral, nasal, topical (including buccal, dermal and sublingual), rectal, vaginal and/or parenteral. The pharmaceutical compositions may conveniently be presented in unit dosage form and may be prepared by any of the methods known in the art of pharmacy. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the patient being treated and the particular mode of administration. The amount of active ingredient which can be used in combination with a carrier material to produce a single dosage form will generally be that amount of the component which produces a therapeutic effect. Typically, expressed as 100%, this amount will range from about 0.01% to about 99% active ingredient, such as from about 0.1% to about 70%, such as from about 1% to about 30%. Irrespective of the chosen route of administration, the compounds of the invention and/or the pharmaceutical compositions of the invention, which may be in the form of pharmaceutically acceptable salts or soluble hydrates, are formulated by conventional methods known to those skilled in the art into pharmaceutically acceptable Accepted Dosage Forms. "Pharmaceutically acceptable salt" refers to a salt that retains the desired biological activity of the parent compound without undesired toxic effects (see, e.g., Berge, S.M. et al., J. Pharm. Sci. 66, 1-19 (1977)). Examples of such salts include salts and bases. Salt addition salts include those derived from nontoxic inorganic acids such as hydrochloric, nitric, similar, sulfuric, hydrobromic, hydroiodic, phosphoric, etc., and from nontoxic organic acids such as aliphatic mono- and dicarboxylic acids, benzene Salts of substituted alkanoic acids, hydroxyalkanoic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. Base-added salts include those derived from alkaline earth metals, such as sodium, potassium, magnesium, calcium, etc., and from non-toxic organic amines, such as N.N'-dibenzylethylenediamine, N-methylglucamine, cloper Salts of nocaine, vitamin B complex, diethanolamine, ethylenediamine, procaine, etc.

药学上可接受的载体包括任何和所有合适的与本发明的化合物兼容的溶剂、分散液、衣料、抗细菌和抗真菌试剂、等压试剂、抗氧化剂和延迟吸收试剂等。Pharmaceutically acceptable carriers include any and all suitable solvents, dispersions, coatings, antibacterial and antifungal agents, isobaric agents, antioxidants and absorption delaying agents, and the like, which are compatible with the compounds of this invention.

用于本发明的药物组合物的合适的水溶性和非水溶性载体的实施例包括水、盐、磷酸缓冲盐、乙醇、葡萄糖、多羟基化合物(例如甘油、丙稀基甘油、聚乙烯甘油等)及其合适的混合物、植物油，例如橄榄油、花生油和芝麻油、羧甲基纤维素胶质溶液、黄芪胶和可注射有机酯，例如乙烷基油酸酯和/或各种缓冲液。其它载体是医学领域已知的。Examples of suitable water-soluble and water-insoluble carriers for the pharmaceutical compositions of the invention include water, saline, phosphate buffered saline, ethanol, dextrose, polyols such as glycerol, acrylglycerin, polyvinylglycerol, etc. ) and suitable mixtures thereof, vegetable oils such as olive oil, peanut oil and sesame oil, carboxymethylcellulose gum solution, tragacanth gum and injectable organic esters such as ethyl oleate and/or various buffers. Other carriers are known in the medical art.

药学上可接受的载体包括无菌水溶液或分散剂和无菌粉剂以制备无菌可注射溶液或分散剂。对药学上活性底物使用这种介质和试剂在本领域是已知的。除了任何与活性化合物不兼容的传统介质或试剂外，其用途在本发明的药物组合物中是可预期的。例如，通过使用包被材料，例如蛋黄素通过维持分散剂所需的颗粒体积，及通过使用表面活性剂来维持适当的流动性。Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the preparation of sterile injectable solutions or dispersions. The use of such media and agents for pharmaceutically active substrates is known in the art. The use of any conventional media or agents incompatible with the active compound is contemplated in the pharmaceutical compositions of the present invention. For example, through the use of coating materials, such as lecithin, to maintain the required particle volume for dispersants, and through the use of surfactants to maintain proper fluidity.

本发明的药物组合物也含有药学上可接受的抗氧化剂，例如(1)水溶性抗氧化剂，例如维生素C酸、半胱氨酸氢氧化物、硫氢酸钠、偏亚硫酸氢钠、亚硫酸钠等；(2)油溶性抗氧化剂，例如抗坏血酸棕榈酸酯、丁基羟基回香醚(BHA)、二丁基羟基甲苯(BHT)、卵磷脂、没食子酸丙酯、维生素E等；及(3)金属螯合剂，例如柠檬酸、乙二胺四乙酸(EDTA)、山梨醇、酒石酸、磷酸等。The pharmaceutical composition of the present invention also contains pharmaceutically acceptable antioxidants, such as (1) water-soluble antioxidants, such as vitamin C acid, cysteine hydroxide, sodium thiohydrogen, sodium metabisulfite, sodium sulfite etc.; (2) Oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyether (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, vitamin E, etc.; and (3 ) Metal chelating agents, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

本发明的药物组合物也可在组分中含有等压试剂，例如糖、多元醇，例如甘露醇、山梨醇、甘油或氯化钠。The pharmaceutical compositions of the present invention may also contain isobaric agents such as sugars, polyalcohols such as mannitol, sorbitol, glycerol or sodium chloride among the components.

药学上可接受的稀释剂包括盐和水溶性缓冲液。Pharmaceutically acceptable diluents include saline and aqueous buffers.

本发明的药物组合物也含有一种或多种适于所选的给药途径的佐剂，例如防腐剂、加湿剂、乳化剂、分散剂、防腐剂或缓冲液，它们可增强药物组合物的货架期或有效性。例如，本发明的化合物可与乳糖、蔗糖、粉剂(例如淀粉)、直链烷酸的纤维素酯、硬脂酸、滑石、硬脂酸镁、氧化镁、磷酸和硫酸的钠、钙盐、阿拉伯树胶、明胶、藻酸钠、聚维酮和/或乙醇。其它佐剂的实施例有QS21、GM-CSF、SRL-172、二盐酸组胺、胸腺卡汀、Tio-TEPA、单磷酰脂A/微细菌组分、明矾、弗氏不完全佐剂、montanide iSA、ribi佐剂系统、TiterMax佐剂、syntex 佐剂配方、免疫刺激复合物(ISCOMs)、gerbu佐剂、CpG脱氧寡核苷酸、脂多糖、和聚肌苷酸:聚胞苷酸。The pharmaceutical compositions of the present invention also contain one or more adjuvants suitable for the chosen route of administration, such as preservatives, wetting agents, emulsifiers, dispersants, preservatives or buffers, which enhance the pharmaceutical composition. shelf life or effectiveness. For example, compounds of the present invention may be formulated with lactose, sucrose, powders such as starch, cellulose esters of linear alkanoic acid, stearic acid, talc, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acids, Gum Arabic, Gelatin, Sodium Alginate, Povidone and/or Alcohol. Examples of other adjuvants are QS21, GM-CSF, SRL-172, histamine dihydrochloride, thymocartin, Tio-TEPA, monophosphoryl lipid A/microbacterial fraction, alum, incomplete Freund's adjuvant, montanide iSA, ribi adjuvant system, TiterMax adjuvant, syntex adjuvant formulation, immunostimulatory complexes (ISCOMs), gerbu adjuvant, CpG deoxyoligonucleotides, lipopolysaccharide, and polyinosinic acid:polycytidylic acid.

抑制微生物的存在可通过灭菌程序和包括各种抗细菌和抗真菌试剂，例如帕拉胶、氯代丁醇、苯酚山梨酸等来保证。另外，可注射药物形式的延时吸收可通过包括延迟吸收的试剂，例如单硬脂酸铝和明胶产生。含本发明的化合物的本发明的药物组合物也可包括合适的盐。任何合适的盐，例如任何合适形式的碱土金属盐(例如缓冲盐)可用于稳定本发明的化合物。合适的盐典型地包括氯化钠、琥珀酸钠、硫酸钠、氯化钾、氯化镁、硫酸镁和氯化钙。在一个技术方案中，铝盐用于稳定本发明的药物组合物中的本发明的化合物，当这种组分对患者给药时，铝盐也作为佐剂。如本发明所述的药物组合物可以多种合适的形式存在。这种形式包括，例如液体、半固体和固体剂量形式，例如液体溶液(例如，可注射和灌液的溶液)、分散液或悬浮液、乳液、微乳液、凝胶、药膏、颗粒粉剂、片剂、药丸、粉剂、脂质体、聚合物和其它纳米颗粒(参见，例如Baek et al.，Methods Enzymol.362，240-9(2003)、Nigavekaret al.，Pharm Res.21(3)，476-83(2004)、微颗粒和栓剂。The inhibition of the presence of microorganisms can be assured by sterilization procedures and the inclusion of various antibacterial and antifungal agents, for example, paragum, chlorobutanol, phenol sorbic acid, and the like. Additionally, prolonged absorption of the injectable pharmaceutical form can be brought about by including agents delaying absorption, for example, aluminum monostearate and gelatin. Pharmaceutical compositions of the invention containing compounds of the invention may also include suitable salts. Any suitable salt, eg, any suitable form of an alkaline earth metal salt (eg, buffer salt) may be used to stabilize the compounds of the invention. Suitable salts typically include sodium chloride, sodium succinate, sodium sulfate, potassium chloride, magnesium chloride, magnesium sulfate and calcium chloride. In one embodiment, aluminum salts are used to stabilize the compounds of the invention in the pharmaceutical compositions of the invention, also as adjuvants when this composition is administered to a patient. The pharmaceutical compositions according to the invention can be in a variety of suitable forms. Such forms include, for example, liquid, semi-solid and solid dosage forms, such as liquid solutions (e.g., injectable and infusion solutions), dispersions or suspensions, emulsions, microemulsions, gels, ointments, granular powders, tablets Dosages, pills, powders, liposomes, polymers and other nanoparticles (see, e.g. Baek et al., Methods Enzymol.362, 240-9 (2003), Nigavekar et al., Pharm Res.21(3), 476 -83 (2004), microgranules and suppositories.

最佳形式依赖于选择的给药方式、组分的性质和治疗用途。例如，配方包括粉剂、泥膏、药膏、凝胶剂、蜡状物、油剂、脂、含(阳离子或阴离子)载体的脂、DNA偶联物、无水吸附剂、水包油和油包水乳液、聚乙二醇(各种分子大小的聚乙二醇)、半固体凝胶和含聚乙二醇的半固体混合物。根据本发明，任何前述形式在治疗脂是合适，只要药物组合物中的活性成分在配制时不失活，而且配方是生理兼容的并可耐受给药途径即可。也参见例如Powell etal.，″Compendium of excipients forparenteral formulations″PDA J Pharm SciTechnol.52，238-311(1998)，此处的引用包括关于药物化学家已知的赋形剂和载体的其它信息。The optimum form depends on the chosen mode of administration, the nature of the ingredients and the therapeutic use. For example, formulations include powders, poultices, salves, gels, waxes, oils, fats, lipids with (cationic or anionic) carriers, DNA conjugates, anhydrous sorbents, oil-in-water and oil-in-water Aqueous emulsions, polyethylene glycols (polyethylene glycols of various molecular sizes), semisolid gels, and semisolid mixtures containing polyethylene glycol. According to the present invention, any of the aforementioned forms is suitable in the treatment of lipids, as long as the active ingredients in the pharmaceutical composition are formulated without inactivation, and the formulation is physiologically compatible and tolerates the route of administration. See also, eg, Powell et al., "Compendium of excipients for parenteral formulations" PDA J Pharm SciTechnol. 52, 238-311 (1998), references therein include additional information on excipients and carriers known to medicinal chemists.

本发明的化合物可用包含化合物免受快速释放，例如以可控释放形式进行释放的载体进行制备，包括植入、皮肤修补和微胶囊递送系统。这种载体可包括明胶、单硬脂酸甘油酯、二硬脂酸甘油酯、生物可降解的生物兼容的多聚物，例如乙烯-乙酸乙烯酯、聚酐、聚羟基乙酸、胶原、聚原酸酯、和聚乳酸单独或与蜡或其它本领域已知的材料一起使用。制备这种配方的方法通常是本领域技术人员已知的，参见例如，Sustainedand ControlledRelease Drug Delivery Systems，J.R.Robinson，ed.，Marcel Dekker，Inc.，New York，1978。为了通过特定给药途径给药本发明的组分，需要将化合物与抑制其失活的化合物包被或与其共同给药。例如，本发明的化合物可在合适载体，例如脂质体或稀释剂中给药患者。脂质体包括油包水CGF乳剂及传统的脂质体(Strejan et al.，J.Neuroimmunol.7，27(1984))。The compounds of the invention can be prepared with a carrier containing the compound against rapid release, eg, in a controlled release form, including implants, skin repairs, and microencapsulated delivery systems. Such carriers may include gelatin, glyceryl monostearate, glyceryl distearate, biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyglycolic acid, Esters, and polylactic acid alone or with waxes or other materials known in the art. Methods for preparing such formulations are generally known to those skilled in the art, see, eg, Sustained and Controlled Release Drug Delivery Systems, J.R. Robinson, ed., Marcel Dekker, Inc., New York, 1978. In order to administer the composition of the present invention by a particular route of administration, it is necessary to coat or co-administer the compound with a compound that inhibits its inactivation. For example, compounds of the invention may be administered to a patient in a suitable carrier, such as liposomes or a diluent. Liposomes include water-in-oil CGF emulsions and conventional liposomes (Strejan et al., J. Neuroimmunol. 7, 27 (1984)).

根据给药途径，活性化合物可包被在材料中，以保护化合物不被酸和其它可能使化合物失活的天然条件的作用。例如，化合物可在合适载体，例如脂质体中给药患者。脂质体包括油包水包油CGF乳剂及传统的脂质体(Strejan et al.，J.Neuroimmunol.7，27(1984))。Depending on the route of administration, the active compound may be coated in materials to protect the compound from acids and other natural conditions that may inactivate the compound. For example, the compounds can be administered to a patient in a suitable carrier, such as liposomes. Liposomes include oil-in-water-in-oil CGF emulsions and conventional liposomes (Strejan et al., J. Neuroimmunol. 7, 27 (1984)).

在一个技术方案中，可配制本发明的化合物，保证其在体内的正确分布。例如，血脑屏障(BBB)可排除多种高亲水性化合物。为了保证本发明的治疗性化合物通过BBB(如果需要)，则它们将配制在例如脂质体中。对制备脂质体的方法，参见例如US 4,522,811、US5,374,548和US 5,399,331。脂质体可含有一个或多个选择性转运到特异细胞或器官中的基团，从而增强靶药的递送(参见例如，V.V.Ranade J.Clin.Pharmacol.29，685(1989))。示例的定位基团包括叶酸或生物素(参见例如US 5,416,016)、甘露糖醇(Umezawa et al.，Biochem.Biophys.Res.Commun.153，1038(1988))、抗体(P.G.Bloeman et al.，FEBSLett.357，140(1995)、M.Owais et al.，Antimicrob.Agents Chemother.39，180(1995))、表面蛋白A受体(Briscoe et al.，Am.J.Physiol.1233.134(1995))、不同物种可含有本发明的药物组合物，及发明的分子的组分，p120(Schreier et al.，J.Biol.Chem.269，9090(1994))，也参见K.Keinanen，M.L.Laukkanen，FEBS Lett.346，123(1994)和JJ.Killion，IJ.Fidler，lmmunomethods 4，273(1994)。In one embodiment, the compounds of the invention can be formulated to ensure their correct distribution in the body. For example, the blood-brain barrier (BBB) excludes many highly hydrophilic compounds. To ensure that the therapeutic compounds of the invention pass the BBB (if desired), they will be formulated, for example, in liposomes. For methods of preparing liposomes see eg US 4,522,811 , US 5,374,548 and US 5,399,331. Liposomes may contain one or more moieties that are selectively transported into specific cells or organs, thereby enhancing delivery of targeted drugs (see, eg, V.V. Ranade J. Clin. Pharmacol. 29, 685 (1989)). Exemplary targeting groups include folic acid or biotin (see e.g. US 5,416,016), mannitol (Umezawa et al., Biochem. Biophys. Res. Commun. 153, 1038 (1988)), antibodies (P.G. Bloeman et al., FEBSLett.357,140(1995), M.Owais et al., Antimicrob.Agents Chemother.39,180(1995)), surface protein A receptor (Briscoe et al., Am.J.Physiol.1233.134(1995) ), different species may contain the pharmaceutical composition of the invention, and components of the molecule of the invention, p120 (Schreier et al., J. Biol. Chem. 269, 9090 (1994)), see also K. Keinanen, M.L. Laukkanen , FEBS Lett. 346, 123 (1994) and JJ. Killion, IJ. Fidler, Immunomethods 4, 273 (1994).

在本发明的一个技术方案中，本发明的化合物配制在脂质体中。在另一个技术方案中，脂质体包括定位基团。在另一个技术方案中，脂质体中的化合物通过快速注射递送到邻近所需部位的位点，例如炎症或感染位点或肿瘤位点。组合物必需是具有易注射性的液体。它必需在制备和储存条件下是稳定的，并可防止微生物，例如细菌和真菌的污染作用。In one technical solution of the present invention, the compound of the present invention is formulated in liposomes. In another embodiment, the liposome includes a targeting group. In another embodiment, the compound in liposomes is delivered by bolus injection to a site adjacent to a desired site, such as a site of inflammation or infection or a tumor site. The composition must be a liquid for easy syringability. It must be stable under the conditions of manufacture and storage and protected against the contaminating action of microorganisms, such as bacteria and fungi.

在一个技术方案中，本发明的化合物可配制为抑制或降低其穿过胎盘的运输。这可通过本领域已知的方法进行，例如通过化合物的PEG化 或使用F(ab′)₂片段。进一步的参考文献是Cunningham-Rundles C et al.，J Immunol Methods.152，177-190(1992)和Landor M.，Ann AllergyAsthma Immunol 74，279-283(1 995)。In one embodiment, the compounds of the invention may be formulated to inhibit or reduce their transport across the placenta. This can be done by methods known in the art, for example by PEGylation of the compound or use of F(ab') ₂ fragments. Further references are Cunningham-Rundles C et al., J Immunol Methods. 152, 177-190 (1992) and Landor M., Ann Allergy Asthma Immunol 74, 279-283 (1995).

用于肠胃外给药的药学上可接受的载体包括无菌的水溶液或分散剂和无菌的粉剂，用于制备无菌的可注射的溶液或分散剂。对药学上的活性底物使用这种介质和试剂在本领域是已知的。除了任何与活性化合物不兼容的传统介质或试剂外，其用途在本发明的药物组合物中是可预期的。Pharmaceutically acceptable carriers for parenteral administration include sterile aqueous solutions or dispersions and sterile powders for the preparation of sterile injectable solutions or dispersions. The use of such media and agents for pharmaceutically active substrates is known in the art. The use of any conventional media or agents incompatible with the active compound is contemplated in the pharmaceutical compositions of the present invention.

附加的活性化合物也可混合在组分中。Additional active compounds can also be mixed into the compositions.

典型地，用于注射的药物组合物必须是无菌的，并在制备和储存条件下是稳定的。化合物可配制为溶液、微乳液、脂质体或其它适于高浓度药物的有序结构。载体可以是水溶性或非水溶性溶剂或含例如水、乙醇、多聚物(例如甘油、丙稀基甘油、聚乙烯甘油等)及其合适的混合物、植物油，例如橄榄油和可注射有机酯，例如乙烷基油酸酯的分散剂。例如，通过使用包被材料，例如蛋黄素通过维持分散剂所需的颗粒体积，及通过使用表面活性剂来维持适当的流动性。在许多情况下，在组分中优选地包括等压试剂，例如糖、多元醇，例如甘油、甘露醇、山梨醇或氯化钠。可注射药物形式的延时吸收可通过在组分中包括可延迟吸收的试剂，例如单硬脂酸盐和明胶产生。无菌注射溶液可通过将活性化合物以所需量混合到所需的具有上述一种或组合成分的合适溶剂中，然后进行微过滤灭菌。通常，通过将活性化合物混合在含基本分散剂和所需的其它成分，例如来自上述成分的无菌载体中而制备。对用无菌粉剂制备无菌注射溶液而言，制备方法的实施例是真空干燥和冻干(冻干法)含活性成分和任何其它所需的，来自前述的无菌过滤溶液的成分。Typically, pharmaceutical compositions for injection must be sterile and stable under the conditions of manufacture and storage. The compounds can be formulated as solutions, microemulsions, liposomes, or other ordered structures suitable to high drug concentration. The carrier can be a water-soluble or non-water-miscible solvent or a solvent containing, for example, water, ethanol, polymers such as glycerol, acrylglycerin, polyvinylglycerol, etc., and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters. , such as ethyl oleate dispersants. For example, through the use of coating materials, such as lecithin, to maintain the required particle volume for dispersants, and through the use of surfactants to maintain proper fluidity. In many cases it is preferred to include isobaric agents in the composition, for example sugars, polyalcohols such as glycerol, mannitol, sorbitol or sodium chloride. Prolonged absorption of the injectable pharmaceutical forms can be brought about by including in the composition agents which delay absorption, for example monostearate and gelatin. Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, followed by microfiltration. In general, the active compound is prepared by incorporating the active compound in a sterile vehicle containing a basic dispersing agent and the required other ingredients, for example, from those mentioned above. For the preparation of sterile injectable solutions from sterile powders, examples of methods of preparation are vacuum drying and freeze-drying (lyophilization) of the active ingredient and any other desired ingredient from previously sterile-filtered solution thereof.

无菌注射溶液可通过将活性化合物以所需量混合到所需的具有上述一种或组合成分的合适溶剂中，然后进行微过滤灭菌。通常，通过将活性化合物混合在含基本分散剂和所需的其它成分，例如来自上述成分的无菌载体中而制备。对用无菌粉剂制备无菌注射溶液而言，制备方法的实施例是真空干燥和冻干(冻干法)含活性成分和任何其它所需的，来自前述的无菌过滤溶液的成分。Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, followed by microfiltration. In general, the active compound is prepared by incorporating the active compound in a sterile vehicle containing a basic dispersing agent and the required other ingredients, for example, from those mentioned above. For the preparation of sterile injectable solutions from sterile powders, examples of methods of preparation are vacuum drying and freeze-drying (lyophilization) of the active ingredient and any other desired ingredient from previously sterile-filtered solution thereof.

本发明的药物组合物可含有一种本发明的化合物或本发明的化合物的组合。因此，在一个技术方案中，本发明的药物组合物包括多个(例 如两个或多个)具有不同作用机制的本发明的化合物的组合，例如一个化合物主要诱导CDC，另一个化合物主要诱导凋亡。A pharmaceutical composition of the invention may contain a single compound of the invention or a combination of compounds of the invention. Therefore, in a technical solution, the pharmaceutical composition of the present invention comprises a combination of a plurality (for example, two or more) of the compounds of the present invention with different mechanisms of action, for example, one compound mainly induces CDC, and the other compound mainly induces apoptosis. Death.

本发明的CD38BPs(包括此处所述的抗CD38抗体、免疫偶联物、双特异性/多特异性分子、组合物和其它衍生物)具有多种体外和体内诊断和治疗用途，可参与由表达CD38的细胞引起的疾病的诊断和治疗。例如，抗体可在例如体外或离体给药所培养的细胞或在例如体内给药受试者，以治疗、抑制和诊断各种疾病。此处所用术语“受试者”是指包括对CD38BP应答的人和非人动物。例如受试者包括具有通过抑制诸如酶活性、信号转导、诱导细胞因子表达、诱导增殖或分化和/或诱导CD38表达细胞的裂解和/或消除或数目降低这样的CD38功能，可校正或减轻疾病的人患者。例如，可用CD38BPs在体内或体外引发一个或多个下述生物学活性：抑制CD38功能(例如酶活性、信号转导、诱导细胞因子表达、诱导增殖或分化和/或诱导裂解)，杀死表达CD38的细胞，在存在人效应细胞时介导表达CD38的细胞的吞噬作用或ADCC，及在存在补体时介导表达CD38的细胞的CDC，或通过凋亡杀死表达CD38的细胞。The CD38BPs of the present invention (including the anti-CD38 antibodies, immunoconjugates, bispecific/multispecific molecules, compositions and other derivatives described herein) have a variety of in vitro and in vivo diagnostic and therapeutic applications, and can participate in the Diagnosis and treatment of diseases caused by cells expressing CD38. For example, antibodies can be administered to cultured cells, eg, in vitro or ex vivo, or to subjects, eg, in vivo, to treat, inhibit, and diagnose various diseases. The term "subject" as used herein is meant to include both human and non-human animals responsive to CD38BP. For example, the subject includes a CD38 function that can be corrected or alleviated by inhibiting CD38 functions such as enzymatic activity, signal transduction, induction of cytokine expression, induction of proliferation or differentiation, and/or induction of lysis and/or depletion or reduction in the number of CD38 expressing cells. disease people patients. For example, CD38BPs can be used to elicit one or more of the following biological activities in vivo or in vitro: inhibition of CD38 function (e.g., enzymatic activity, signal transduction, induction of cytokine expression, induction of proliferation or differentiation, and/or induction of lysis), killing of expressed CD38-expressing cells, mediate phagocytosis or ADCC of CD38-expressing cells in the presence of human effector cells, and mediate CDC of CD38-expressing cells in the presence of complement, or kill CD38-expressing cells by apoptosis.

在存在补体时，也可使用任何含具有补体结合位点，例如结合补体的来源于IgG1、-2或-3或IgM的一部分的本发明的CD38BPs的组分。在一个技术方案中，离体治疗含具有本发明的CD38BP的靶细胞和合适的效应细胞的细胞群可通过加入补体或含补体的血清而进行补充。被本发明的CD38BP包被的靶细胞的吞噬作用或裂解可通过补体蛋白的结合而增强。在一个技术方案中，包被本发明的CD38BPs的靶细胞也可被补体裂解。在一个技术方案中，本发明的CD38BPs不激活补体。In the presence of complement, any composition comprising CD38BPs of the invention having a complement binding site, eg, a portion derived from IgGl, -2 or -3 or IgM, that binds complement may also be used. In one embodiment, ex vivo treatment of a population of cells comprising target cells bearing a CD38BP of the invention and suitable effector cells may be supplemented by addition of complement or complement-containing serum. Phagocytosis or lysis of target cells coated with CD38BP of the present invention can be enhanced by binding of complement proteins. In one technical solution, target cells coated with CD38BPs of the present invention can also be lysed by complement. In one technical solution, the CD38BPs of the present invention do not activate complement.

本发明的CD38BPs也可与补体一起给药。因此，含血清或补体的CD38BPs也在本发明的范围中。在这些组分中，补体通过例如偶联而紧邻CD38BPs，或适于进行同时给药。选择性地，CD38BPs和补体或血清可单独给药。The CD38BPs of the invention can also be administered with complement. Therefore, CD38BPs containing serum or complement are also within the scope of the present invention. In these components, complement is in close proximity to CD38BPs, eg by conjugation, or is suitable for simultaneous administration. Alternatively, CD38BPs and complement or serum can be administered alone.

本发明的CD38BPs也用于针对表达FcγR或CD38的细胞，例如用于标记这类细胞。对这种用途而言，CD38BP可与能被检测的分子连接。因此，本发明提供离体或在体外定位表达Fc受体，例如FcγR或CD38的细胞的方法。可检测标记可以是例如放射性同位素、荧光化合物、酶或酶辅因子。CD38BPs of the invention are also useful for targeting cells expressing FcyR or CD38, for example for labeling such cells. For this use, CD38BP can be linked to a molecule that can be detected. Accordingly, the present invention provides methods for localizing ex vivo or in vitro cells expressing Fc receptors, such as FcyR or CD38. A detectable label can be, for example, a radioisotope, a fluorescent compound, an enzyme or an enzyme cofactor.

靶特异的效应细胞，例如与本发明的CD38BP连接的效应细胞也可用作治疗性试剂。定位的效应细胞可以是人淋巴细胞，例如巨噬细胞、嗜中性粒细胞或单核细胞。其它细胞包括嗜曙红细胞、自然杀伤细胞和其它含IgG或IgA受体的细胞。如果需要，效应细胞可从被治疗的患者中获得。靶特异的效应细胞可在生理上可接受的溶液中作为悬浮液进行给药。给药的细胞数目可在10⁸到10⁹，但可根据治疗目的而改变。通常使用可在靶细胞，例如表达CD38的肿瘤细胞获得定位，并通过例如吞噬作用或裂解而有效杀死细胞的足够的量。Target-specific effector cells, eg, effector cells linked to a CD38BP of the invention may also be used as therapeutic agents. The targeted effector cells may be human lymphocytes such as macrophages, neutrophils or monocytes. Other cells include eosinophils, natural killer cells, and other IgG or IgA receptor-containing cells. Effector cells can be obtained from the patient being treated, if desired. Target-specific effector cells can be administered as a suspension in a physiologically acceptable solution. The number of cells administered can range from 10 ⁸ to 10 ⁹ , but can vary depending on the therapeutic purpose. Sufficient amounts are generally used to achieve localization in target cells, eg, CD38-expressing tumor cells, and to effectively kill the cells, eg, by phagocytosis or lysis.

用靶特异的效应细胞的治疗可联合其它清楚靶细胞的技术来进行。例如，使用本发明的CD38BPs和/或针对这些组分的效应细胞的抗肿瘤治疗可与化疗联合使用。另外，联合免疫治疗可用于指导两种不同的细胞毒素效应物群针对肿瘤细胞抑制。例如与抗FcγRI或抗CD3连接的CD38BP可与IgG或IgA受体特异结合试剂联合使用。本发明的双特异性和多特异性分子也可用于例如通过在细胞表面加帽和删除受体来调控效应细胞上的FcαR或FcγR水平。抗Fc受体的混合物也可用于该目的。Therapy with target-specific effector cells can be performed in conjunction with other techniques for elucidating target cells. For example, antitumor therapy using CD38BPs of the invention and/or effector cells directed against these components can be used in combination with chemotherapy. Additionally, combination immunotherapy can be used to direct two distinct cytotoxic effector populations against tumor cell suppression. For example, CD38BP linked to anti-FcyRI or anti-CD3 can be used in combination with IgG or IgA receptor specific binding reagents. Bispecific and multispecific molecules of the invention can also be used to modulate FcαR or FcγR levels on effector cells, eg, by capping and deleting receptors on the cell surface. Cocktails of anti-Fc receptors can also be used for this purpose.

在一个技术方案中，本发明提供在样品中检测CD38抗原存在或测定CD38抗原的量的方法，包括将样品和对照样品与特异结合CD38的CD38BP在可在CD38BP或其部分和CD38间形成复合物的条件下接触。然后检测复合物的形成，其中与对照样品相比较，样品间形成复合物的差异表明在样品中存在CD38抗原。检测免疫检测方法的实施例包括但不限定于ELISA、RIA、FACS检测、等离子共振检测、色谱检测、组织免疫组织化学、Western印迹和/或免疫沉淀。In one technical solution, the present invention provides a method for detecting the presence of CD38 antigen or measuring the amount of CD38 antigen in a sample, comprising combining the sample and the control sample with CD38BP that specifically binds CD38 to form a complex between CD38BP or its part and CD38 contact under conditions. Complex formation is then detected, wherein a difference in complex formation between samples compared to a control sample indicates the presence of CD38 antigen in the sample. Examples of detection immunoassay methods include, but are not limited to, ELISA, RIA, FACS detection, plasmon resonance detection, chromatographic detection, tissue immunohistochemistry, Western blotting, and/or immunoprecipitation.

在一个技术方案中，本发明的CD38BPs可用于检测CD38的循环水平或在其膜表面含CD38的细胞水平，然后将该水平与特定的疾病症状相关联。选择性地，CD38BPs可用于清除或与表达CD38的细胞相互作用，从而表明这些细胞是疾病的重要介导物。也可通过将样品和对照样品与抗CD38抗体在适于在抗体和CD38形成复合物的条件下接触。检测抗体和CD38形成的任何复合物，并将样品和对照进行比较。本发明的CD38BPs最初可在体外检测治疗和诊断用途相关的结合活性。例如，可用流式细胞仪检测CD38BPs。另外，可检测CD38BPs在引发至少一种效应细胞介导的效应细胞活性方面的活性。例如，可检测本发明 的抗CD38抗体引发CDC和/或凋亡的能力。检测CDC、同型黏附、分子成簇或凋亡的流程在本领域是已知的。In one technical solution, the CD38BPs of the present invention can be used to detect the circulating level of CD38 or the level of cells containing CD38 on their membrane surface, and then correlate the level with a specific disease symptom. Alternatively, CD38BPs can be used to clear or interact with CD38-expressing cells, thereby suggesting that these cells are important mediators of disease. It can also be achieved by contacting the sample and control samples with an anti-CD38 antibody under conditions suitable for complex formation of the antibody and CD38. Any complexes formed by the antibody and CD38 were detected and samples were compared to controls. The CD38BPs of the present invention can initially be tested in vitro for binding activity relevant to therapeutic and diagnostic uses. For example, CD38BPs can be detected by flow cytometry. Additionally, the activity of CD38BPs in eliciting at least one effector cell-mediated effector cell activity can be detected. For example, anti-CD38 antibodies of the invention can be tested for their ability to elicit CDC and/or apoptosis. Protocols for detecting CDC, homotypic adhesion, molecular clustering or apoptosis are known in the art.

在一个技术方案中，本发明提供在体内或体外检测并定量CD38表达细胞的量的方法。该方法包括(i)给药受试者偶联可检测标记的本发明的CD38BP；(ii)将受试者暴露在检测所述可检测标记中，以鉴定含CD38表达细胞的区域。In one technical solution, the present invention provides a method for detecting and quantifying the amount of CD38 expressing cells in vivo or in vitro. The method comprises (i) administering to a subject a CD38BP of the invention coupled to a detectable marker; (ii) exposing the subject to detection of the detectable marker to identify regions containing CD38 expressing cells.

在一个技术方案中，本发明的免疫偶联物可通过以这种靶化合物作为本发明的免疫偶联物中的治疗性基团，而将化合物(例如治疗性试剂、标记、细胞毒素、免疫抑制剂等)定位到具有结合到表面的CD38的细胞上。In a technical solution, the immunoconjugate of the present invention can be used to convert the compound (such as therapeutic agent, label, cytotoxin, immune inhibitors, etc.) to cells with CD38 bound to the surface.

在一个技术方案中，本发明也提供离体或在体外定位表达CD38的细胞的方法(例如，用可检测标记，例如放射性同位素、荧光化合物、酶或酶辅因子)。In one embodiment, the invention also provides methods for localizing CD38-expressing cells ex vivo or in vitro (eg, with a detectable label such as a radioisotope, fluorescent compound, enzyme or enzyme cofactor).

在一个技术方案中，本发明提供通过给药本发明的免疫毒素而杀死具有结合到表面的CD38的细胞。In one embodiment, the invention provides killing of cells having CD38 bound to the surface by administering an immunotoxin of the invention.

本发明提供在患者中治疗或预防由CD38表达细胞参与的疾病，该方法包括对需要治疗的患者给药治疗有效量的本发明的CD38BP。这种CD38BPs用于抑制CD38诱导的与疾病相关的活性或消除或降低CD38表达细胞的数目。The present invention provides treating or preventing diseases involving CD38 expressing cells in patients, the method comprising administering a therapeutically effective amount of CD38BP of the present invention to the patient in need of treatment. Such CD38BPs are used to inhibit CD38-induced disease-related activity or to eliminate or reduce the number of CD38-expressing cells.

这种方法包括对患者给药有效量的本发明的CD38BP组分以治疗或预防疾病。CD38BP组分可单独给药或与其它治疗性药物，例如本文其它地方所述的与CD38BP联合或协同作用的试剂来治疗或预防CD38表达细胞参与的疾病。选择性地，免疫偶联物可用于通过专门针对CD38的细胞毒素或放射性毒素而杀死在其表面表达CD38的细胞。Such methods include administering to a patient an effective amount of a CD38BP component of the invention to treat or prevent a disease. The CD38BP component can be administered alone or in combination with other therapeutic agents, such as agents described elsewhere herein that combine or act synergistically with CD38BP, to treat or prevent diseases in which CD38-expressing cells are involved. Alternatively, immunoconjugates can be used to kill cells expressing CD38 on their surface by cytotoxins or radiotoxins specific for CD38.

在本发明的一个技术方案中，表达CD38的细胞参与的疾病可以是肿瘤性疾病，例如疾病的特征在于存在表达CD38的肿瘤细胞，例如B细胞淋巴瘤、浆细胞恶性瘤、T/NK细胞淋巴瘤和骨髓恶性瘤。In one technical solution of the present invention, the disease in which CD38-expressing cells participate can be a neoplastic disease, for example, a disease characterized by the presence of CD38-expressing tumor cells, such as B-cell lymphoma, plasma cell malignancy, T/NK cell lymphoma tumors and myeloid malignancies.

这种肿瘤性疾病的实施例包括B细胞淋巴瘤/白血病，包括前体B细胞成淋巴细胞白血病/淋巴瘤和B细胞非霍奇金淋巴瘤；急性早幼粒细胞白血病急性成淋巴细胞白血病和成熟B细胞肿瘤，例如B细胞慢性淋巴细胞性白血病(CLL)/小淋巴细胞性白血病(SLL)、B细胞急性淋巴细胞性白血病、B细胞前淋巴细胞白血病、淋巴浆细胞样淋巴瘤、 套细胞淋巴瘤(MCL)、包括低级、中级和高级FL在内的滤泡性淋巴瘤(FL)、皮肤滤泡中心淋巴瘤、边缘区B细胞淋巴瘤(MALT型、结与脾型)、毛细胞白血病、弥漫性大B细胞淋巴瘤、Burkitt淋巴瘤、浆细胞瘤、浆细胞骨髓瘤、浆细胞白血病、移植后淋巴增生性疾病、Waldenstrom巨球蛋白血症、浆细胞白血病和间变性大细胞淋巴瘤(ALCL)。Examples of such neoplastic diseases include B-cell lymphoma/leukemia, including precursor B-cell lymphoblastic leukemia/lymphoma and B-cell non-Hodgkin's lymphoma; acute promyelocytic leukemia, acute lymphoblastic leukemia, and Mature B-cell neoplasms such as B-cell chronic lymphocytic leukemia (CLL)/small lymphocytic leukemia (SLL), B-cell acute lymphoblastic leukemia, B-cell prolymphocytic leukemia, lymphoplasmacytoid lymphoma, mantle cell Lymphoma (MCL), follicular lymphoma (FL) including low-, intermediate-, and high-grade FL, cutaneous follicle center lymphoma, marginal zone B-cell lymphoma (MALT, nodal, and splenic), hair cell Leukemia, diffuse large B-cell lymphoma, Burkitt lymphoma, plasmacytoma, plasma cell myeloma, plasma cell leukemia, post-transplantation lymphoproliferative disorder, Waldenstrom's macroglobulinemia, plasma cell leukemia, and anaplastic large cell lymphoma tumor (ALCL).

在一个技术方案中，由表达CD38的细胞参与的疾病是多发性骨髓瘤。In one embodiment, the disease involving cells expressing CD38 is multiple myeloma.

B细胞非霍奇金淋巴瘤的实施例有淋巴瘤样肉芽肿病、原发性渗出性淋巴瘤、血管内大B细胞淋巴瘤、纵隔大B细胞淋巴瘤、重链疾病(包括γ、μ和α疾病)、由免疫抑制试剂诱导的淋巴瘤，例如环孢霉素诱导的淋巴瘤和甲氨蝶呤诱导的淋巴瘤。Examples of B-cell non-Hodgkin's lymphomas are lymphomatoid granulomatous disease, primary effusion lymphoma, intravascular large B-cell lymphoma, mediastinal large B-cell lymphoma, heavy chain disease (including gamma, mu and alpha diseases), lymphomas induced by immunosuppressive agents such as cyclosporine-induced lymphomas and methotrexate-induced lymphomas.

在本发明的一个技术方案中，由表达CD38的细胞参与的疾病是霍奇金淋巴瘤。In one technical solution of the present invention, the disease involving CD38-expressing cells is Hodgkin's lymphoma.

由表达CD38的细胞参与的疾病的实施例可以是来源于T和NK细胞的恶性瘤，包括：成熟T细胞和NK细胞瘤，包括T细胞前淋巴细胞白血病、T细胞大颗粒淋巴细胞白血病、侵袭性NK细胞白血病、成人T细胞白血病/淋巴瘤、结外NK-T细胞淋巴瘤、鼻型、肠病型T细胞淋巴瘤、肝脾性T细胞淋巴瘤、皮下脂膜炎样T细胞淋巴瘤、母细胞性NK细胞淋巴瘤、蕈样霉菌病/Sezary症候群、原发性皮肤CD30阳性T细胞淋巴增生性疾病(原发性皮肤间变性大细胞淋巴瘤C-ALCL、淋巴瘤样丘疹病、交界性病变)、血管免疫母细胞性T细胞淋巴瘤、非特指型外周T细胞淋巴瘤和退行性大细胞淋巴瘤。Examples of diseases involving CD38-expressing cells may be malignancies derived from T and NK cells, including: mature T-cell and NK-cell neoplasms, including T-cell prolymphocytic leukemia, T-cell large granular lymphocytic leukemia, invasive NK-cell leukemia, adult T-cell leukemia/lymphoma, extranodal NK-T-cell lymphoma, nasal type, enteropathic T-cell lymphoma, hepatosplenic T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, Blastoid NK-cell lymphoma, mycosis fungoides/Sezary syndrome, primary cutaneous CD30-positive T-cell lymphoproliferative disorders (primary cutaneous anaplastic large cell lymphoma C-ALCL, lymphomatoid papulosis, junctional angioimmunoblastic T-cell lymphoma, unspecified peripheral T-cell lymphoma, and anaplastic large cell lymphoma.

来源于骨髓细胞的恶性瘤的实施例包括急性骨髓白血病，包括急性早幼粒细胞性白血病、和包括慢性粒细胞白血病在内的慢性髓细胞白血病。Examples of malignant tumors derived from myeloid cells include acute myeloid leukemia, including acute promyelocytic leukemia, and chronic myeloid leukemia including chronic myeloid leukemia.

在本发明的一个技术方案中，由表达CD38的细胞参与的疾病是由CD38表达B细胞、浆细胞、单核细胞和T细胞参与的免疫性疾病。In one technical solution of the present invention, the disease involving CD38-expressing cells is an immune disease involving CD38-expressing B cells, plasma cells, monocytes and T cells.

由CD38表达B细胞、浆细胞、单核细胞和T细胞参与的免疫性疾病的实施例包括诸如牛皮癣、关节银屑病、皮炎、系统性硬皮病及硬化症这样的自身免疫失调、炎症性肠病(IBD)、Crohn病、溃疡性结肠炎、呼吸窘迫综合征、脑膜炎、脑炎、葡萄膜炎、肾小球肾炎、湿疹、 哮喘、动脉硬化、白细胞粘附缺陷病、多发性硬化症、Raynaud症候群、Sjogren症候群、青少年糖尿病、Reiter病、Behcet病、免疫复合物性肾炎、IgA肾病、IgM多发性神经病、诸如急性特发性血小板减少性紫癜及慢性特发性血小板减少性紫癜这样的免疫介导的血小板减少症状、溶血性贫血、重症肌无力、狼疮性肾炎、系统性红斑狼疮、风湿关节炎(RA)、异位性皮炎、天疱疮、Graves病、桥本甲状腺炎、Wegener肉芽肿、Omenn症候群、慢性肾功能衰竭、急性传染性单核细胞增多征、多发性硬化症、HIV和疱疹病毒相关的疾病。进一步实施例包括严重急性呼吸综合征和脉络视网膜炎(choreoretinitis)。另外，也包括其它疾病和症状，例如那些由诸如伊波病毒(EBV)这样的病毒感染B细胞引起或介导的疾病。Examples of immune diseases involving CD38 expressing B cells, plasma cells, monocytes and T cells include autoimmune disorders such as psoriasis, psoriasis joints, dermatitis, systemic scleroderma and sclerosis, inflammatory Intestinal disease (IBD), Crohn's disease, ulcerative colitis, respiratory distress syndrome, meningitis, encephalitis, uveitis, glomerulonephritis, eczema, asthma, arteriosclerosis, leukocyte adhesion deficiency disease, multiple sclerosis syndrome, Raynaud syndrome, Sjogren syndrome, juvenile diabetes, Reiter disease, Behcet disease, immune complex nephritis, IgA nephropathy, IgM polyneuropathy, such as acute idiopathic thrombocytopenic purpura and chronic idiopathic thrombocytopenic purpura Immune-mediated thrombocytopenia, hemolytic anemia, myasthenia gravis, lupus nephritis, systemic lupus erythematosus, rheumatoid arthritis (RA), atopic dermatitis, pemphigus, Graves disease, Hashimoto's thyroiditis, Wegener Granuloma, Omenn's syndrome, chronic renal failure, acute infectious mononucleosis, multiple sclerosis, HIV, and herpes virus-related diseases. Further examples include severe acute respiratory syndrome and choreoretinitis. In addition, other diseases and conditions are also included, such as those caused or mediated by infection of B cells by viruses such as Epstein-Barr virus (EBV).

在一个技术方案中，由表达CD38的细胞参与的疾病是风湿性关节炎。In one embodiment, the disease involving CD38-expressing cells is rheumatoid arthritis.

其中自身抗体和/或过量B和T淋巴细胞活性显著并可根据本发明进行治疗的炎症性、免疫性和/或自免疫性疾病的进一步实施例包括以下部分：Further examples of inflammatory, immune and/or autoimmune diseases in which autoantibodies and/or excess B and T lymphocyte activity are significant and which can be treated according to the invention include the following:

血管炎和其它血管疾病，譬如显微镜下多血管炎、Churg-Strauss综合征、及其它ANCA相关性血管炎、结节性多动脉炎、原发性冷球蛋白血症血管炎、皮肤白细胞分裂性血管炎、川崎病、大动脉炎、巨细胞动脉炎、过敏性紫癜肾炎、原发性或独立的脑脉管炎、结节性红斑、血栓闭塞性脉管炎、血栓性血小板减少性紫癜(包括溶血性尿毒综合征)、以及包括皮肤白细胞碎裂性血管炎(例如第二期的B型肝炎、C型肝炎、Waldenstrom巨球蛋白血症、B细胞瘤样变、类风湿性关节炎、Sjogren综合征、或系统性红斑狼疮)在内的继发性血管炎；进一步的实施例有结节性红斑、变应性血管炎、脂膜炎、复发性结节性非化脓性脂膜炎、紫癜hyperglobulinaemica、及Buerger病；皮肤病，譬如接触性皮炎、线状IgA皮肤病、白癜风、坏疽性脓皮病、获得性大疱性表皮松解症、寻常性天疱疮(包括疤痕性类天疱疮和大疱性类天疱疮)、斑秃(包括普秃和全秃)、疱疹样皮炎、多形红斑、与慢性自身免疫性荨麻疹(包括血管神经性水肿和荨麻疹性血管炎)；Vasculitis and other vascular diseases such as microscopic polyangiitis, Churg-Strauss syndrome, and other ANCA-associated vasculitis, polyarteritis nodosa, primary cryoglobulinemia vasculitis, cutaneous leukocytosis Vasculitis, Kawasaki disease, Takayasu arteritis, giant cell arteritis, Henoch-Schonlein purpura nephritis, primary or isolated cerebral vasculitis, erythema nodosum, thromboangiitis obliterans, thrombotic thrombocytopenic purpura (including hemolytic uremic syndrome), and including cutaneous leukocytoclastic vasculitis (eg, second stage hepatitis B, hepatitis C, Waldenstrom's macroglobulinemia, B-cell neoplasia, rheumatoid arthritis, Sjogren syndrome, or systemic lupus erythematosus); further examples are erythema nodosum, allergic vasculitis, panniculitis, recurrent nodular nonsuppurative panniculitis, Purpura hyperglobulinaemica, and Buerger's disease; skin diseases such as contact dermatitis, linear IgA dermatosis, vitiligo, pyoderma gangrenosum, epidermolysis bullosa acquired, pemphigus vulgaris (including cicatricial pemphigoid herpes and bullous pemphigoid), alopecia areata (including alopecia generalis and alopecia totalis), dermatitis herpetiformis, erythema multiforme, and chronic autoimmune urticaria (including angioedema and urticarial vasculitis) ;

免疫介导血细胞减少症，譬如自身免疫性中性粒细胞减少症和单纯红细胞再生障碍性贫血；Immune-mediated cytopenias such as autoimmune neutropenia and pure red cell aplasia;

结缔组织病，譬如中枢神经系统狼疮、盘状红斑狼疮、CREST综合征、混合性结缔组织病、多肌炎/皮肌炎、包涵体肌炎、继发性淀粉样变性、I型及II型冷沉球蛋白血症、纤维肌痛、磷脂抗体综合征、继发性血友病、复发性多发软骨炎、结节病、僵人综合征和风湿热；进一步的实施例有嗜曙红细胞筋膜炎；Connective tissue diseases such as central nervous system lupus, discoid lupus erythematosus, CREST syndrome, mixed connective tissue disease, polymyositis/dermatomyositis, inclusion body myositis, secondary amyloidosis, types I and II Cryoglobulinemia, fibromyalgia, phospholipid antibody syndrome, secondary hemophilia, relapsing polychondritis, sarcoidosis, stiff man syndrome, and rheumatic fever; further examples are eosinophilia Meningitis;

关节炎，譬如强直性脊柱炎、幼年性慢性关节炎、成人Still病及SAPHO综合征；进一步的实施例有骶骨关节炎、反应性关节炎、Still病和痛风病；Arthritis, such as ankylosing spondylitis, juvenile chronic arthritis, adult Still's disease and SAPHO syndrome; further examples are sacral arthritis, reactive arthritis, Still's disease and gout;

血液病、譬如再生障碍性贫血、原发性溶血性贫血(包括冷凝集素综合征)、对CLL或者系统性红斑狼疮的继发性溶血性贫血；POEMS综合征、恶性贫血及Waldemstróm′spurpura hyperglobulinaemica；进一步的实施例有粒性白血球缺乏症、自身免疫性中性粒细胞减少症、Franklin病、Seligmann病、γ重链疾病、对胸腺瘤和淋巴瘤的继发性副癌综合征、及因子VIII抑制物形成；Blood disorders such as aplastic anemia, primary hemolytic anemia (including cold agglutinin syndrome), secondary hemolytic anemia to CLL or systemic lupus erythematosus; POEMS syndrome, pernicious anemia, and Waldemstróm'spurpura hyperglobulinaemica ; further examples are agranulocytosis, autoimmune neutropenia, Franklin disease, Seligmann disease, gamma heavy chain disease, secondary paraneoplastic syndromes to thymoma and lymphoma, and factor VIII inhibitor formation;

内分泌病、譬如多内分泌症和阿狄森病；进一步的实施例有自身免疫性低血糖症、自身免疫性甲状腺功能低下、胰岛素自身免疫综合征、德奎尔万甲状腺炎、及胰岛素受体抗体介导的胰岛素抵抗；Endocrinopathies, such as polyendocrinology and Addison's disease; further examples are autoimmune hypoglycemia, autoimmune hypothyroidism, insulin autoimmune syndrome, de Quervan's thyroiditis, and insulin receptor antibodies mediated insulin resistance;

肝-胃肠疾病，譬如乳糜泻、Whipple病、原发性胆汁性硬变、慢性活动性肝炎、及原发性硬化性胆管炎；进一步的实施例是自身免疫性胃炎：Hepato-gastrointestinal diseases such as celiac disease, Whipple's disease, primary biliary cirrhosis, chronic active hepatitis, and primary sclerosing cholangitis; a further example is autoimmune gastritis:

肾病，譬如快速进行性肾小球肾炎、后链球菌肾炎、肺出血-肾炎综合征、膜性肾小球肾炎和冷球蛋白肾炎；进一步的实施例是微小病变疾病；Kidney diseases such as rapidly progressive glomerulonephritis, posterior streptococcal nephritis, pulmonary hemorrhage-nephritic syndrome, membranous glomerulonephritis and cryoglobulin nephritis; further examples are minimal change disease;

神经障碍、譬如自身免疫性神经病、多数性单神经炎、Lambert-Eaton肌无力综合征、Sydenham舞蹈病、脊髓痨及Guillain-Barre综合征；进一步的实施例有脊髓病/热带痉挛性瘫痪、重症肌无力、急性炎症性脱髓鞘性多发性神经炎及慢性炎症性脱髓鞘性多发性神经炎；多发性硬化症；Nervous disorders such as autoimmune neuropathy, mononeuritis majority, Lambert-Eaton myasthenic syndrome, Sydenham chorea, tabes and Guillain-Barre syndrome; further examples are myelopathy/tropical spastic paralysis, severe Myasthenia, acute inflammatory demyelinating polyneuritis, and chronic inflammatory demyelinating polyneuritis; multiple sclerosis;

心脏及肺部疾病、譬如COPD、纤维性肺泡炎、闭塞性细支气管炎、过敏性肺部 菌病、囊性纤维变性、Loffler综合征、心肌炎及心包炎；进一步的实施例有过敏性肺炎和继发于肺癌的肿瘤伴随综合征；Heart and lung diseases, such as COPD, fibrous alveolitis, bronchiolitis obliterans, allergic lung mycosis, cystic fibrosis, Loffler syndrome, myocarditis and pericarditis; further examples are hypersensitivity pneumonitis and tumor-associated syndrome secondary to lung cancer;

变态反应性疾病、譬如支气管性气喘和高lgE综合征；进一步的实 施例是一时性黑蒙；Allergic diseases, such as bronchial asthma and hyper IgE syndrome; a further example is amaurosis;

眼科疾病，譬如先天性脉络膜视网膜炎；Eye diseases, such as congenital chorioretinitis;

感染性疾病，譬如细小病毒B感染(包括hands-and-socks syndrome在内)；Infectious diseases, such as parvovirus B infection (including hands-and-socks syndrome);

妇产科疾病，譬如反复流产、反复胎儿损失及宫内发育迟缓；进一步的实施例是继发于妇科肿瘤的肿瘤伴随综合征；Gynecological disorders, such as recurrent miscarriage, recurrent fetal loss and intrauterine growth retardation; a further example is tumor-associated syndrome secondary to gynecological tumors;

男性生殖系统疾病，譬如继发于睾丸肿瘤的肿瘤伴随综合征；Diseases of the male reproductive system, such as tumor-associated syndrome secondary to testicular tumors;

以及由于移植引起的疾病，譬如同种异体移植物与异种移植物排斥及移植物抗宿主病。And diseases caused by transplantation, such as allograft and xenograft rejection and graft-versus-host disease.

也可预防性给药抗体以降低发生诸如上述肿瘤性疾病这样的癌症的危险性，延迟在这种癌症进程中的发作，和/或在清除这类癌症后降低复发危险性。这在根据其它生物学因素已知存在肿瘤但难于对其定位的患者中是特别有用的。Antibodies may also be administered prophylactically to reduce the risk of developing, delay the onset of such cancers in the course of their progression, and/or reduce the risk of recurrence after such cancers have been cleared, such as the neoplastic diseases described above. This is particularly useful in patients where tumors are known to be present based on other biological factors but are difficult to locate.

本发明的组分包括“治疗有效量的”或“预防有效量的”CD38BP。“治疗有效量的”是指在需要的时间内可达到所需治疗结果的有效剂量。CD38BP的治疗有效量可根据诸如个体的疾病状态、年龄、性别和体重，及CD38BP在个体中引发所需的应答等因素而改变。治疗有效量也是其中抗体或抗体部分的任何有毒或有害作用超过治疗有利作用的量。“预防有效量”是指在需要的时间内可达到所需预防结果(例如降低发展疾病的可能性、降低疾病的强度和扩散性、在疾病即将来临时增加存活的可能性、延迟疾病症状的发作等)的有效剂量。典型地，由于用于患者的预防剂量在疾病早期阶段之前，因此预防性有效量将低于治疗性有效剂量。对肿瘤治疗的“治疗性有效量”也可通过其稳定疾病进展的能力来确定。化合物抑制癌症的能力可在动物模式系统中评估其在人肿瘤中的有效性。选择性地，组分的这一特性可通过技术人员已知的体外检测法检测化合物抑制细胞生长或诱导凋亡的能力来评估。治疗性化合物的治疗有效量可在患者中减小肿瘤体积或减轻症状。本领域技术人员可根据诸如患者的体重、患者病症的严重性和所选择的特定化合物或给药途径来确定该剂量。The composition of the present invention includes a "therapeutically effective amount" or "prophylactically effective amount" of CD38BP. A "therapeutically effective amount" refers to an amount effective to achieve the desired therapeutic result, for the period of time required. A therapeutically effective amount of CD38BP can vary depending on factors such as the disease state, age, sex and weight of the individual, and the CD38BP eliciting the desired response in the individual. A therapeutically effective amount is also one in which any toxic or detrimental effects of the antibody or antibody portion outweigh the therapeutically beneficial effects. A "prophylactically effective amount" means an amount that achieves the desired prophylactic result (e.g., reduces the likelihood of developing disease, reduces the intensity and spread of disease, increases the likelihood of survival when disease is imminent, delays symptoms of disease) for the time needed. seizures, etc.) effective dose. Typically, prophylactically effective doses will be lower than therapeutically effective doses since prophylactic doses are administered to patients prior to the early stages of the disease. A "therapeutically effective amount" for tumor treatment can also be determined by its ability to stabilize disease progression. The ability of a compound to inhibit cancer can be evaluated in animal model systems for its effectiveness in human tumors. Alternatively, this property of a component can be assessed for the ability of the compound to inhibit cell growth or induce apoptosis by in vitro assays known to the skilled artisan. A therapeutically effective amount of a therapeutic compound reduces tumor volume or alleviates symptoms in a patient. The dosage can be determined by one skilled in the art based on factors such as the patient's weight, the severity of the patient's condition and the particular compound or route of administration chosen.

风湿性关节炎的“治疗有效量”可在患者中产生至少ACR₂₀初步疗效判定标准，例如至少ACR₅₀初步疗效判定标准，例如至少ARC₇₀初步疗效判定标准。ACR₂₀初步疗效判定标准定义为：在压痛关节计数 (TJC)提高≥20％，并在以下5项评估中提高≥20％：患者疼痛评估(VAS)1、患者整体评估(VAS)、医生整体评估(VAS)、患者自测无力(HAQ)急性阶段反应物(CRP或ESR)。ACR₅₀和ACR₇₀以相同方式定义为分别≥50％和≥70％的提高。对于进一步的细节，参见Felsonet al.，in American College of Rheumatology PreliminaryDefinition ofImprovement in Rheumatoid Arthriffs；Arthritis Rheumatism 38，727-735(1995)。A "therapeutically effective amount" for rheumatoid arthritis produces at least an ACR ₂₀ preliminary response criterion, eg, at least an ACR ₅₀ preliminary response criterion, eg, at least an ARC ₇₀ preliminary response criterion, in the patient. ACR ₂₀ preliminary efficacy criteria are defined as: ≥ 20% improvement in tender joint count (TJC), and ≥ 20% improvement in the following 5 assessments: Patient Pain Assessment (VAS)1, Patient Overall Assessment (VAS), Physician Overall Assessment assessment (VAS), patient self-assessment of weakness (HAQ), acute phase reactants (CRP or ESR). ACR ₅₀ and ACR ₇₀ were defined in the same way as improvements of >50% and >70%, respectively. For further details see Felson et al., in American College of Rheumatology Preliminary Definition of Improvement in Rheumatoid Arthriffs; Arthritis Rheumatism 38, 727-735 (1995).

选择性地，风湿性关节炎的治疗有效量可通过由EULAR定义的DAS(疾病活动度评分)，包括DAS28和/或DAS56来测定。Alternatively, the therapeutically effective amount for rheumatoid arthritis can be determined by DAS (Disease Activity Score) defined by EULAR, including DAS28 and/or DAS56.

调整剂量以提供最适的所需应答(例如，治疗性应答)。例如，可进行单点注射给药，也可随时间进行几次分开的剂量给药，或根据治疗情况的紧急性按比例减小或增加剂量。肠胃外组分可配制成易于给药的剂量单位和通用剂量形式。此处所用剂量单位形式是指适于被治疗患者的个别的适于作为单一剂量的形式；每个单位含预定量的活性化合物以在与所需药物载体联合时产生所需的治疗效果。本发明的剂量单位形式的规格由和直接依赖于(a)活性化合物的特定性质和需达到的特定治疗效果，及(b)配制方面本身的限制，例如用于治疗的活性化合物在个体中的灵敏度而确定。本发明的CD38BPs的有效剂量和剂量形式根据被治疗疾病或症状而由本领域技术人员来确定。本发明的化合物的治疗有效剂量的示例的非限制性范围是约0.1-100mg/kg、譬如约0.1-50mg/kg、例如约0.1-20mg/kg、譬如约0.1-10mg/kg、例如约0.5、譬如约0.3、约1或约3mg/kg。Dosage is adjusted to provide the optimum desired response (eg, a therapeutic response). For example, administered as a single injection, several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. Parenteral compositions can be formulated in dosage units and general dosage forms for ease of administration. Dosage unit form as used herein refers to individual forms suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specifications for the dosage unit forms of the invention consist of and are directly dependent on (a) the particular properties of the active compound and the particular therapeutic effect to be achieved, and (b) limitations inherent in formulation, such as the amount of active compound used for treatment in an individual. Determined by sensitivity. Effective doses and dosage forms of CD38BPs of the present invention can be determined by those skilled in the art according to the disease or condition to be treated. Exemplary non-limiting ranges of therapeutically effective doses of compounds of the invention are about 0.1-100 mg/kg, such as about 0.1-50 mg/kg, such as about 0.1-20 mg/kg, such as about 0.1-10 mg/kg, such as about 0.5 , such as about 0.3, about 1 or about 3 mg/kg.

本领域的医生或兽医可很容易地确定并给药有效剂量的所需药物组合物。例如，医生或兽医开始可使用比达到所需治疗效果所需的剂量低的药物组合物中所用的本发明的CD38BPs，然后逐渐增加剂量以达到所需效果。通常，本发明的组分的合适每日剂量是指化合物的剂量是有效产生治疗性效果的最低剂量。这种有效剂量通常根据上述因素所确定。可进行静脉内、肌肉内、腹膜内或皮下给药，例如对靶位点最近处给药。如果需要，药物组合物的有效每日剂量可在一天的合适间隔时间内分别进行2、3、4、5、6或更多次亚剂量给药，选择性地，以单位剂量形式进行给药。但对本发明的化合物可进行单独给药，优选地如上述药物组合物一样给药化合物。A physician or veterinarian skilled in the art can readily determine and administer an effective dose of the desired pharmaceutical composition. For example, a physician or veterinarian may initially use lower doses of the CD38BPs used in the pharmaceutical composition than are required to achieve the desired therapeutic effect, and then gradually increase the dose to achieve the desired effect. In general, suitable daily dosages of the components of the invention mean that the dosage of the compound is the lowest dosage effective to produce a therapeutic effect. Such an effective dose will generally be determined based on the factors mentioned above. Administration can be intravenous, intramuscular, intraperitoneal or subcutaneous, for example proximal to the target site. If desired, the effective daily dose of the pharmaceutical composition may be administered in 2, 3, 4, 5, 6 or more sub-doses at suitable intervals throughout the day, optionally in unit dose form . However, the compound of the present invention can be administered alone, preferably the compound is administered as the above-mentioned pharmaceutical composition.

在一个技术方案中，本发明的CD38BPs可以每周剂量为从10到500mg/m²，譬如从200到400mg/m²通过灌液进行给药。这种给药可重复例如1到8次，譬如3到5次。可在2到24小时，例如从2到12小时的时间内通过连续灌液进行给药。In a technical solution, the CD38BPs of the present invention can be administered through irrigation at a weekly dose of from 10 to 500 mg/m ² , such as from 200 to 400 mg/m ² . This administration may be repeated, for example, 1 to 8 times, such as 3 to 5 times. Administration may be by continuous infusion over a period of 2 to 24 hours, eg, from 2 to 12 hours.

在一个技术方案中，本发明的CD38BPs可在较长的时间内，例如24小时以上，通过缓慢的连续灌液进行给药，以降低毒副作用。In a technical solution, the CD38BPs of the present invention can be administered through slow continuous infusion over a long period of time, such as over 24 hours, so as to reduce toxic and side effects.

在一个技术方案中，本发明的CD38BPs可以每周剂量为从250mg到2000mg，譬如例如300mg、500mg、700mg、1000mg、1500mg或2000mg进行8次以上，例如从4到6次给药。可在一段时间内，例如从2到24小时，譬如从2到12小时通过连续灌液进行给药。如果需要，这种形式可重复1次或多次，例如，在6个月或12个月后重复进行。可在给药后，通过例如提取生物样品，并用针对本发明的CD38BPs的抗原结合区域的抗独特型抗体测定血液中本发明的化合物的量来确定或调整剂量。In a technical solution, the CD38BPs of the present invention can be administered at a weekly dose of from 250 mg to 2000 mg, such as 300 mg, 500 mg, 700 mg, 1000 mg, 1500 mg or 2000 mg, for more than 8 times, such as from 4 to 6 times. Administration may be by continuous infusion over a period of time, for example from 2 to 24 hours, such as from 2 to 12 hours. This form can be repeated 1 or more times if desired, for example, after 6 or 12 months. The dose can be determined or adjusted after administration by, for example, extracting a biological sample and measuring the amount of the compound of the present invention in the blood with an anti-idiotypic antibody directed against the antigen-binding region of the CD38BPs of the present invention.

在一个技术方案中，本发明的CD38BPs可通过维持治疗进行给药，譬如例如，1周1次进行6个月或更长时间。In one technical solution, the CD38BPs of the present invention can be administered by maintenance therapy, such as, for example, once a week for 6 months or longer.

在一个技术方案中，本发明的CD38BPs可通过包括本发明的CD38BP灌液，然后用偶联放射性同位素的本发明的CD38BP灌液进行给药。给药可在例如7到9天后重复进行。In one technical solution, the CD38BPs of the present invention can be administered by infusion containing the CD38BP of the present invention, followed by infusion of the CD38BP of the present invention coupled with a radioactive isotope. Dosing can be repeated, eg, after 7 to 9 days.

作为非限制性实施例，如本发明所述的治疗可以每日本发明的化合物的剂量为0.1-100mg/kg来提供，例如每天0.5、0.9、1.0、1.1、1.5、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、40、45、50、60、70、80、90或100mg/kg，在初始治疗后，在至少第1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、 28、29、30、31、32、33、34、35、36、37、38、39或40天，或选择性地，至少第1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19或20周，或其组合，用单个或分开剂量每隔24、12、8、6、4或2小时或其任何组合给药一次。As a non-limiting example, treatment according to the invention may be provided at a dose of 0.1-100 mg/kg of the compound of the invention per day, for example 0.5, 0.9, 1.0, 1.1, 1.5, 2, 3, 4, 5 , 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 , 40, 45, 50, 60, 70, 80, 90, or 100 mg/kg, after initial treatment, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 , 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37 , 38, 39 or 40 days, or alternatively, at least the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th , 19 or 20 weeks, or a combination thereof, administered in single or divided doses every 24, 12, 8, 6, 4, or 2 hours or any combination thereof.

本发明的药物组合物也以联合治疗的形式给药，例如，与其他被治疗疾病或症状相关的治疗性药剂联合使用。这种给药可以是同时的、单独的或连续的。对同时给药而言，药剂可以合适的1个组分或单独的组 分进行给药。因此，本发明提供治疗上述表达CD38的细胞参与的疾病的方法，方法包括与以下所述的一个或多个其他治疗性药剂联合给药本发明的CD38BP。The pharmaceutical compositions of the invention are also administered in combination therapy, for example, with other therapeutic agents associated with the disease or condition being treated. Such administration may be simultaneous, separate or sequential. For simultaneous administration, the agents may be administered as a suitable component or as separate components. Therefore, the present invention provides a method for treating the above-mentioned diseases in which CD38-expressing cells are involved, the method comprising administering CD38BP of the present invention in combination with one or more other therapeutic agents described below.

本发明也提供本发明的CD38BP在制备与至少一种针对上述表达CD38的细胞参与的化疗试剂一起给药的药物组合物方面的用途。The present invention also provides the use of the CD38BP of the present invention in the preparation of a pharmaceutical composition administered together with at least one chemotherapeutic agent targeting the above CD38-expressing cells.

在一个技术方案中，联合治疗可包括将本发明的组分与至少一种化疗试剂，至少一种抗炎症试剂或至少一种免疫抑制剂和/或免疫调节剂联合给药。In one technical solution, combination therapy may comprise administering the composition of the present invention in combination with at least one chemotherapeutic agent, at least one anti-inflammatory agent or at least one immunosuppressant and/or immunomodulator.

在一个技术方案中，本发明提供在患者中治疗由表达CD38的细胞参与的疾病的方法，该方法包括给药所需患者治疗有效剂量的本发明的CD3 8BP和至少一种化疗试剂。In one embodiment, the present invention provides a method of treating a disease involving CD38 expressing cells in a patient, the method comprising administering to the patient in need thereof a therapeutically effective amount of a CD3 8BP of the invention and at least one chemotherapeutic agent.

在一个技术方案中，本发明提供治疗多发性骨髓瘤的方法，该方法包括给药所需患者治疗有效剂量的本发明的CD38BP和至少一种化疗试剂。In one embodiment, the present invention provides a method of treating multiple myeloma comprising administering to a patient in need thereof a therapeutically effective amount of a CD38BP of the present invention and at least one chemotherapeutic agent.

在一个技术方案中，本发明提供本发明的CD38BP在制备与至少一种化疗试剂联合给药以治疗多发性骨髓瘤的药物组合物方面的用途。In one technical solution, the present invention provides the use of the CD38BP of the present invention in the preparation of a pharmaceutical composition administered in combination with at least one chemotherapeutic agent for the treatment of multiple myeloma.

在一个技术方案中，这种化疗试剂选自抗代谢物，譬如甲氨蝶呤、6-巯基嘌呤、6-硫鸟嘌呤、阿拉伯树胶酸、氟达拉滨、5-氟尿嘧啶、氨烯咪胺、羟基脲、天冬酰胺酶、吉西他滨、克拉屈滨和类似试剂。In one embodiment, the chemotherapeutic agent is selected from antimetabolites such as methotrexate, 6-mercaptopurine, 6-thioguanine, arabic acid, fludarabine, 5-fluorouracil, carbamide , hydroxyurea, asparaginase, gemcitabine, cladribine and similar agents.

在一个技术方案中，这种化疗试剂选自烷基化试剂，例如二氯甲基二乙胺、thioepa、瘤可宁、美法仑、双氯乙基亚硝脲(BSNU)、氮芥(CCNU)、环磷酰胺、白消安、二溴甘露醇、链脲霉素、达卡巴嗪(DTIC)、甲基苄肼、丝裂霉素C、施铂锭及其它诸如卡铂这样的铂衍生物和类似试剂。In a technical scheme, this chemotherapeutic agent is selected from the group consisting of alkylating agents, such as dichloromethyldiethylamine, thioepa, leucoridine, melphalan, bischloroethylnitrosourea (BSNU), nitrogen mustard ( CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, dacarbazine (DTIC), procarbazine, mitomycin C, platinum, and other platinum agents such as carboplatin Derivatives and similar reagents.

在一个技术方案中，这种化疗试剂选自抗生素，例如更生霉素(以前称为放射菌素)、博来霉素、正定霉素(以前称为道诺霉素)、阿霉素、去甲氧正定霉素、光神霉素、丝裂霉素、米托蒽醌、光辉霉素、安曲霉素(AMC)和类似试剂。In one embodiment, the chemotherapeutic agent is selected from antibiotics such as dactinomycin (formerly known as actinomycin), bleomycin, daunomycin (formerly known as daunorubicin), doxorubicin, Methodomycin, mithramycin, mitomycin, mitoxantrone, mitoxantrone, antramycin (AMC) and similar agents.

在一个技术方案中，这种化疗试剂选自抗有丝分裂试剂，例如紫杉烷、例如紫杉萜和太平洋紫杉醇，长春花碱、例如长春地辛、长春新碱、长春碱和长春瑞滨。In one embodiment, the chemotherapeutic agent is selected from antimitotic agents such as taxanes such as docetaxel and paclitaxel, vinblastines such as vindesine, vincristine, vinblastine and vinorelbine.

在一个技术方案中，这种化疗试剂选自拓扑异构酶抑制剂，例如托 泊替康。In one embodiment, the chemotherapeutic agent is selected from topoisomerase inhibitors, such as topotecan.

在一个技术方案中，这种化疗试剂选自生长因子抑制剂，例如ErbB1(EGFR)(例如吉非替尼(Iressa)、西妥昔单抗(Erbitux)、埃罗替尼(Tarceva)、HuMax-EGFr(2F8，在WO 2002/100348)描述)和类似试剂)的抑制剂，ErbB2(Her2/neu)(例如曲妥单抗(Herceptin)和类似试剂)的抑制剂和类似试剂。In a technical solution, the chemotherapeutic agent is selected from growth factor inhibitors, such as ErbB1 (EGFR) (such as gefitinib (Iressa ), Cetuximab (Erbitux ), Erlotinib (Tarceva ), HuMax-EGFr (2F8, described in WO 2002/100348)) and similar agents), inhibitors of ErbB2 (Her2/neu) (such as trastuzumab (Herceptin ) and similar reagents) inhibitors and similar reagents.

在一个技术方案中，这种生长因子抑制剂是法呢基转移酶抑制剂，例如SCH-66336和R115777。In one embodiment, the growth factor inhibitor is a farnesyl transferase inhibitor, such as SCH-66336 and R115777.

在一个技术方案中，这种生长因子抑制剂是血管内皮生长因子(VEGF)抑制剂，例如贝伐单抗(Avastin)。In one technical solution, the growth factor inhibitor is a vascular endothelial growth factor (VEGF) inhibitor, such as bevacizumab (Avastin ).

在一个技术方案中，这种化疗试剂是酪氨酸激酶抑制剂，例如伊马替尼(Glivec，Gleevec STI571)、拉帕替尼泊、PTK787/ZK222584和类似试剂。In one embodiment, such chemotherapeutic agents are tyrosine kinase inhibitors such as imatinib (Glivec, Gleevec STI571 ), lapatinib, PTK787/ZK222584 and similar agents.

在一个技术方案中，这种化疗试剂是组氨酸去乙酰化酶抑制剂。这种组氨酸去乙酰化酶抑制剂的实施例包括基于异羟肟酸的杂合极性化合物，譬如SAHA(辛二酰苯胺异羟肟酸)。In one embodiment, the chemotherapeutic agent is a histidine sirtuin inhibitor. Examples of such histidine sirtuin inhibitors include hydroxamic acid-based hybrid polar compounds such as SAHA (suberoylanilide hydroxamic acid).

在一个技术方案中，这种化疗试剂是诸如SCIO-469这样的P38aMAP激酶抑制剂。In one embodiment, the chemotherapeutic agent is a P38aMAP kinase inhibitor such as SCIO-469.

在一个技术方案中，本发明提供在患者中治疗由表达CD38的细胞参与的疾病的方法，该方法包括给药所需患者治疗有效剂量的本发明的CD38BP和至少一种血管生成、新血管生成和/或其它血管化抑制剂。In one technical solution, the present invention provides a method for treating a disease involving CD38-expressing cells in a patient, the method comprising administering a therapeutically effective dose of CD38BP of the present invention and at least one of angiogenesis, neovascularization, and and/or other vascularization inhibitors.

在一个技术方案中，本发明提供治疗多发性骨髓瘤的方法，该方法包括给药所需患者治疗有效剂量的本发明的CD38BP和至少一种血管生成、新血管生成和/或其它血管化抑制剂。In one technical solution, the present invention provides a method of treating multiple myeloma comprising administering to a patient in need thereof a therapeutically effective dose of CD38BP of the present invention and at least one angiogenesis, neovascularization and/or other vascularization inhibitory agent.

在一个技术方案中，本发明提供本发明的CD38BP在制备与至少一种血管生成、新血管生成和/或其它血管化抑制剂联合给药以治疗多发性骨髓瘤的药物组合物方面的用途。In one technical solution, the present invention provides the use of CD38BP of the present invention in the preparation of a pharmaceutical composition administered in combination with at least one angiogenesis, neovascularization and/or other vascularization inhibitors to treat multiple myeloma.

这种血管生成抑制剂的实施例是尿激酶抑制剂、基质金属蛋白酶抑制剂(例如马马司他、新伐司他、BAY12-9566、AG3340、BMS-275291和类似试剂)、内皮细胞迁移和增殖抑制剂(例如TNP-470、角鲨胺、2-甲氧雌二醇、考布他汀、内皮他丁、血管他丁、青霉胺、SCH66336(Schering-Plough Corp，Madison，NJ)、R115777(Janssen Pharmaceutica，Inc，Titusville，NJ)和类似试剂)、血管生成因子拮抗剂(例如譬如ZD6474、SU6668、抗血管生成试剂的抗体和/或其受体(例如VEGF、bFGF和血管生成素-1)、沙利度胺(Thalomid)、沙利度胺类似物(譬如CC-5013(来那度胺，ReVLimid^TM)和CC4047(Actimid^TM)、Sugen 5416、SU5402、抗血管生成因子的核酶(譬如angiozyme)、干扰素α(譬如干扰素α2a)、苏拉明及类似试剂)、VEGF-R激酶抑制剂和其它抗血管生成因子的酪氨酸激酶抑制剂(譬如SU011248)、内皮特异整联蛋白/存活信号传导抑制剂(譬如牡荆碱及类似试剂)、铜拮抗剂/螯合剂(譬如四硫钼酸盐、卡普多普瑞尔和类似试剂)、羧基氨基咪唑(CAI)、ABT-627、CM101、白介素-12(IL-12)、IM862、PNU145156E及抑制血管生成的核苷酸分子(譬如反义-VEGF-cDNA、编码血管生成抑制素的cDNA、编码p53的cDNA和编码缺陷型VEGF受体2的cDNA)和类似试剂。Examples of such angiogenesis inhibitors are urokinase inhibitors, matrix metalloproteinase inhibitors (e.g. marimastat, neovalastat, BAY12-9566, AG3340, BMS-275291 and similar agents), endothelial cell migration and Proliferation inhibitors (eg, TNP-470, squalamine, 2-methoxyestradiol, combretastatin, endostatin, angiostatin, penicillamine, SCH66336 (Schering-Plough Corp, Madison, NJ), R115777 (Janssen Pharmaceutica, Inc, Titusville, NJ) and similar agents), angiogenic factor antagonists (such as ZD6474, SU6668, antibodies to anti-angiogenic agents and/or their receptors (such as VEGF, bFGF, and angiopoietin-1 ), Thalidomide (Thalomid ), thalidomide analogs (such as CC-5013 (lenalidomide, ReVLimid ^TM ) and CC4047 (Actimid ^TM ), Sugen 5416, SU5402, ribozymes of anti-angiogenic factors (such as angiozyme), interferon alpha ( such as interferon alpha 2a), suramin and similar agents), VEGF-R kinase inhibitors and tyrosine kinase inhibitors of other anti-angiogenic factors (such as SU011248), endothelial-specific integrin/survival signaling inhibitors ( such as viteline and similar agents), copper antagonists/chelators (such as tetrathiomolybdate, captopril and similar agents), carboxyaminoimidazole (CAI), ABT-627, CM101, interleukin-12 (IL-12), IM862, PNU145156E, and nucleotide molecules that inhibit angiogenesis (such as antisense-VEGF-cDNA, cDNA encoding angiomotin, cDNA encoding p53, and cDNA encoding defective VEGF receptor 2) and similar reagents.

其它这种血管生成、新血管生成和/或其它血管化的抑制剂的实施例是抗血管生成的苷磷脂衍生物和相关分子(例如，heperinase III)、替莫唑胺、NK4、巨噬细胞转移抑制因子(MIF)、环氧合酶-2抑制物、缺氧诱导因子1抑制物、抗血管生成大豆异黄酮、奥替普拉、烟曲霉素及其类似物、生长抑素类似物、戊聚糖多硫酸酯、替可加兰钠、达肝素、肿瘤抑素、凝血酶致敏蛋白、NM-3、考布他汀、血管生成抑制素、阿伐他汀、抗其它相关靶标的抗体(例如抗α-v/β-3整联蛋白和抗激肽抑制素(kininostatin)mAbs和类似试剂。Examples of other such inhibitors of angiogenesis, neovascularization and/or other vascularization are anti-angiogenic glycoside phospholipid derivatives and related molecules (e.g., heperinase III), temozolomide, NK4, macrophage metastasis suppressor (MIF), cyclooxygenase-2 inhibitors, hypoxia-inducible factor 1 inhibitors, anti-angiogenic soy isoflavones, oltipraz, fumagillin and its analogs, somatostatin analogs, pentameric Glycopolysulfate, tecogalan sodium, dalteparin, tumorstatin, thrombin sensitized protein, NM-3, combretastatin, angiostatin, atorvastatin, antibodies against other relevant targets (such as anti- α-v/β-3 integrin and anti-kininostatin mAbs and similar reagents.

在一个技术方案中，本发明提供本发明的CD38BP在制备与沙利度胺(Thalomid)、沙利度胺类似物(譬如CC-5013(来那度胺、ReVlimid^TM)和/或CC4047(Actimid^TM)联合给药方面的用途。In one technical scheme, the present invention provides CD38BP of the present invention in preparation with Thalidomide (Thalomid ), thalidomide analogs (such as CC-5013 (Lenalidomide, ReVlimid ^TM ) and/or CC4047 (Actimid ^TM ) in combination administration.

在一个技术方案中，本发明提供本发明的CD38BP在制备与沙利度胺联合给药的药物组合物方面的用途。In one technical solution, the present invention provides the use of the CD38BP of the present invention in the preparation of a pharmaceutical composition administered in combination with thalidomide.

在一个技术方案中，本发明提供本发明的CD38BP在制备与抗CD20抗体，譬如利妥昔单抗(Rituxan，Mabthera)、如WO 2004/035607所述的人单克隆抗CD20抗体，譬如11B8、2F2或7D8联合给药的药物组合物方面的用途。In one technical scheme, the present invention provides that CD38BP of the present invention is prepared with anti-CD20 antibody, such as rituximab (Rituxan , Mabthera ), the use of a human monoclonal anti-CD20 antibody as described in WO 2004/035607, such as 11B8, 2F2 or 7D8, in a pharmaceutical composition administered in combination.

在一个技术方案中，与本发明的CD38BPs联合使用以治疗上述疾病的治疗性试剂是蛋白酶抑制剂，例如硼替佐米(Velcade)。In one technical solution, the therapeutic agent used in combination with CD38BPs of the present invention to treat the above-mentioned diseases is a protease inhibitor, such as bortezomib (Velcade ).

在一个技术方案中，与本发明的CD38BPs联合使用以治疗上述疾病的治疗性试剂是皮质类固醇，例如强的松、脱氢皮质甾醇、地塞米松等。In one technical solution, the therapeutic agent used in combination with the CD38BPs of the present invention to treat the above-mentioned diseases is a corticosteroid, such as prednisone, prednisone, prednisone, dexamethasone and the like.

在一个技术方案中，与本发明的CD38BPs联合使用以治疗上述疾病的治疗性试剂是皮质类固醇，例如强的松、脱氢皮质甾醇、地塞米松等。In one technical solution, the therapeutic agent used in combination with the CD38BPs of the present invention to treat the above-mentioned diseases is a corticosteroid, such as prednisone, prednisone, prednisone, dexamethasone and the like.

在一个技术方案中，与本发明的CD38BPs联合使用以治疗上述疾病的治疗性试剂是抗癌免疫原，例如癌抗原/肿瘤相关的抗原(例如内皮细胞黏附分子(EpCAM/TACSTDI)、粘液素1(MUC1)、癌胚抗原(CEA)、肿瘤相关的糖蛋白72(TAG-72)、gplOO、Melan-A、MART-1、KDR、RCAS1、MDA7、癌相关病毒疫苗(例如，人乳头瘤病毒疫苗)、肿瘤来源的热休克蛋白和类似试剂。此处所述多种其它合适的癌抗原/肿瘤相关抗原和本领域已知的类似分子也可选择性地用于这种技术方案中。抗癌免疫原性蛋白也包括抗独特型“疫苗”，例如BEC2抗独特型抗体、米妥莫单抗、CeaVac和相关的抗独特型抗体，针对MG7抗体的抗独特型抗体及其它抗癌抗独特型抗体(参见例如，Birebent et al.，Vaccine.21.(15)，1601-12(2003)、Li etal.，Chin Med J(Engl).114(9)，962-6(2001)、Schmitt et al.，Hybridoma.13(5)，389-96(1994)、Maloney et al.，Hybridoma.4(3)，191-209(1985)、Raychardhuri et al.，JImmunol.137(5)，1743-9(1986)、Pohl et al.，lnt J Cancer.50(6)，958-67(1992)、Bohlen et al.，Cytokines MoI Ther.2(4)，231-8(1996)和Maruyama，J ImmunolMethods.264(1-2).121-33(2002))。这种抗独特型Abs可选择性地与作为合成(典型地惰性)分子载体、蛋白(例如钥孔血蓝蛋白(KLH)(参见例如，Ochi et al.，Eur J Immunol.17(11)，1645-8(1987))、或细胞(例如红血细胞，参见例如Wi et al.，J ImmunolMethods.122(2)，227-34(1989))载体偶联。In one technical solution, the therapeutic agent used in combination with CD38BPs of the present invention to treat the above-mentioned diseases is an anti-cancer immunogen, such as cancer antigen/tumor-associated antigen (such as endothelial cell adhesion molecule (EpCAM/TACSTDI), mucin 1 (MUC1), carcinoembryonic antigen (CEA), tumor-associated glycoprotein 72 (TAG-72), gplOO, Melan-A, MART-1, KDR, RCAS1, MDA7, cancer-associated virus vaccines (e.g., human papillomavirus Vaccines), tumor-derived heat shock proteins, and similar agents. A variety of other suitable cancer antigens/tumor-associated antigens described herein and similar molecules known in the art can optionally be used in this technical scheme. Cancer immunogenic proteins also include anti-idiotypic "vaccines" such as BEC2 anti-idiotypic antibodies, Mitumomab, CeaVac and related anti-idiotypic antibodies, anti-idiotypic antibodies against MG7 antibodies and other anti-idiotype antibodies against cancer Type antibodies (see for example, Birebent et al., Vaccine. 21. (15), 1601-12 (2003), Li et al., Chin Med J (Engl). 114 (9), 962-6 (2001), Schmitt et al., Hybridoma.13(5), 389-96(1994), Maloney et al., Hybridoma.4(3), 191-209(1985), Raychardhuri et al., JImmunol.137(5), 1743 -9(1986), Pohl et al., lnt J Cancer.50(6), 958-67(1992), Bohlen et al., Cytokines MoI Ther.2(4), 231-8(1996) and Maruyama, J ImmunolMethods.264(1-2).121-33(2002)). Such anti-idiotypic Abs can be selectively combined with synthetic (typically inert) molecular carriers, proteins (such as keyhole limpet hemocyanin (KLH) (see for example, Ochi et al., Eur J Immunol.17 (11), 1645-8 (1987)), or cell (for example red blood cell, see for example Wi et al., J ImmunolMethods.122 (2), 227- 34 (1989)) carrier coupling.

在一个技术方案中，与本发明的CD38BPs联合使用以治疗上述疾病的治疗性试剂是二磷酸盐。可能合适的二磷酸盐的实施例是帕米膦酸盐(Aredia)、唑来膦酸(Zometa)、氯膦酸盐(Bonefos)、利塞膦酸盐(Actonel)、伊班膦酸盐(Boniva)、依替膦酸盐(Didronel)、阿仑膦酸盐(Fosamax)、替鲁膦酸盐(Skelid)、伊卡膦酸盐(Yamanouchi Pharmaceutical)和米诺膦酸盐(YM529、 Yamanouchi)。In one technical solution, the therapeutic agent used in combination with the CD38BPs of the present invention to treat the above-mentioned diseases is a bisphosphonate. An example of a potentially suitable bisphosphonate is pamidronate (Aredia ), zoledronic acid (Zometa ), clodronate (Bonefos ), risedronate (Actonel ), ibandronate (Boniva ), etidronate (Didronel ), alendronate (Fosamax ), tiludronate (Skelid ), icadronate (Yamanouchi Pharmaceutical), and minodronate (YM529, Yamanouchi).

在一个技术方案中，与本发明的CD38BPs联合使用以治疗上述疾病的治疗性试剂是集落刺激因子。合适的集落刺激因子的实施例是粒细胞集落刺激因子(G-CSF)，例如非格司亭(Neupogen)和聚乙二醇化非格司亭(Neulasta)，和粒细胞巨噬细胞集落刺激因子(GM-CSF)，例如沙格司亭(Leukine)。In one technical solution, the therapeutic agent used in combination with the CD38BPs of the present invention to treat the above-mentioned diseases is a colony-stimulating factor. An example of a suitable colony stimulating factor is granulocyte colony stimulating factor (G-CSF), such as filgrastim (Neupogen ) and pegfilgrastim (Neulasta ), and granulocyte-macrophage colony-stimulating factor (GM-CSF), such as sargragrastim (Leukine ).

在一个技术方案中，与本发明的CD38BPs联合使用以治疗上述疾病的治疗性试剂是红血球生成试剂。合适的红血球生成试剂的实施例是红细胞生成素(EPO)，例如红细胞生成素α(例如Procrit、Epogen 和Eprex)和红细胞生成素β(例如NeoRecormon)和红血球生成刺激蛋白(例如Aranesp)。In one technical solution, the therapeutic agent used in combination with the CD38BPs of the present invention to treat the above diseases is an erythropoietic agent. An example of a suitable erythropoietic agent is erythropoietin (EPO), such as erythropoietin alpha (e.g. Procrit 、Epogen and Eprex ) and erythropoietin β (such as NeoRecormon ) and erythropoiesis-stimulating proteins (such as Aranesp ).

在一个技术方案中，与本发明的CD38BPs联合使用以治疗上述疾病的治疗性试剂是抗癌细胞因子或其组合。合适的细胞因子和生长因子的实施例包括IFNγ、IL-2、IL-4、IL-6、IL-7、IL-10、IL-12、IL-13、IL-15、IL-18、IL-23、IL-24、IL-27、IL-28a、IL-28b、IL-29、KGF、IFNα(例如INFα2b)、IFNβ3、GM-CSF、CD40L、FItS配体、干细胞因子、安西司亭和TNFα。合适的趋化因子包括来自人CXC和C-C趋化因子家族的Glu-Leu-Arg(ELR)-阴性趋化因子，例如IP-10、MCP-3、MIG和SDF-1α。合适的细胞因子包括细胞因子衍生物、细胞因子变异体、细胞因子片段和细胞因子融合蛋白。In one technical solution, the therapeutic agent used in combination with the CD38BPs of the present invention to treat the above-mentioned diseases is an anti-cancer cell factor or a combination thereof. Examples of suitable cytokines and growth factors include IFNγ, IL-2, IL-4, IL-6, IL-7, IL-10, IL-12, IL-13, IL-15, IL-18, IL -23, IL-24, IL-27, IL-28a, IL-28b, IL-29, KGF, IFNα (eg INFα2b), IFNβ3, GM-CSF, CD40L, FItS ligand, stem cell factor, anxigrastim and TNFα. Suitable chemokines include Glu-Leu-Arg (ELR)-negative chemokines from the human CXC and C-C chemokine families, such as IP-10, MCP-3, MIG and SDF-la. Suitable cytokines include cytokine derivatives, cytokine variants, cytokine fragments and cytokine fusion proteins.

此处这些和其它天然肽编码核酸参与的方法或用途可选择性地或另外通过如US5,968,502、US 6,063,630和US 6,187,305及EP 0505500中所述的“基因激活”和同源重组基因上调技术来进行。Methods or uses in which these and other native peptide-encoding nucleic acids involve can alternatively or additionally be achieved by "gene activation" and homologous recombination gene up-regulation techniques as described in US 5,968,502, US 6,063,630 and US 6,187,305 and EP 0505500 conduct.

在一个技术方案中，与本发明的CD38BPs联合使用以治疗上述疾病的治疗性试剂是例如增强或抑制Fcα或Fcγ受体的表达或活性的调控试剂。适于该用途的试剂的实施例包括白介素-1(IL-1)、白介素-2(IL-2)、白介素-6(IL-6)、粒细胞集落刺激因子(G-CSF)，例如非格司亭(Neupogen)和聚乙二醇化非格司亭(Neulasta)，及粒细胞巨噬细胞集落刺激因子(GM-CSF)，例如沙格司亭(Leukine)，干扰素-γ(IFN-γ)和肿瘤坏死因子(TNF)。In one technical solution, the therapeutic agent used in combination with the CD38BPs of the present invention to treat the above-mentioned diseases is, for example, a regulatory agent that enhances or inhibits the expression or activity of Fcα or Fcγ receptors. Examples of agents suitable for this use include interleukin-1 (IL-1), interleukin-2 (IL-2), interleukin-6 (IL-6), granulocyte colony stimulating factor (G-CSF), e.g. Grastim (Neupogen ) and pegfilgrastim (Neulasta ), and granulocyte-macrophage colony-stimulating factor (GM-CSF), such as sargragrastim (Leukine ), interferon-γ (IFN-γ) and tumor necrosis factor (TNF).

在一个技术方案中，与本发明的CD38BPs联合使用以治疗上述疾病的治疗性试剂是细胞循环控制/凋亡调控子(或“调控试剂”)。细胞 循环控制/凋亡调控子包括以下分子(i)定位和调控细胞循环控制/凋亡调控子，例如cdc-25(例如NSC 663284)，(ii)过度刺激细胞循环的细胞周期蛋白依赖的激酶(例如黄酮类抗肿瘤药flavopiridol(L868275，HMR1275)、7-hydroxystaurosporine(UCN-01，KW-2401)和roscovitine(R-roscovitine，CYC202))，及(iii)端粒酶调控子(例如B1BR1532、SOT-095、GRN163就例如US 6,440,735和US 6,713,055中描述的组分)。干扰凋亡途径的分子的非限制性实施例包括TNF-相关的凋亡诱导配体(TRAI L)/凋亡-2配体(Apo-2L)、诱导NF-κB阻塞从而抑制IL-6产生的试剂，激活TRAIL受体的抗体、IFNs1反义Bcl-2和As₂O₃ (三氧化二砷，Trisenox)。In one technical solution, the therapeutic agent used in combination with the CD38BPs of the present invention to treat the above-mentioned diseases is a cell cycle control/apoptosis regulator (or "regulatory agent"). Cell cycle control/apoptosis regulators include molecules that (i) localize and regulate cell cycle control/apoptosis regulators such as cdc-25 (eg NSC 663284), (ii) cyclin-dependent kinases that overstimulate the cell cycle (such as flavonoid antineoplastic drugs flavopiridol (L868275, HMR1275), 7-hydroxystaurosporine (UCN-01, KW-2401) and roscovitine (R-roscovitine, CYC202)), and (iii) telomerase regulators (such as B1BR1532, SOT-095, GRN163 are eg components described in US 6,440,735 and US 6,713,055). Non-limiting examples of molecules that interfere with the apoptotic pathway include TNF-related apoptosis-inducing ligand (TRAIL)/apoptosis-2 ligand (Apo-2L), induction of NF-κB blockage thereby inhibiting IL-6 production Reagents, antibodies that activate TRAIL receptors, IFNs1 antisense Bcl-2 and As ₂ O ₃ (arsenic trioxide, Trisenox ).

在一个技术方案中，与本发明的CD38BPs联合使用以治疗上述疾病的治疗性试剂是激素调控试剂，例如用于抗雄激素和抗雌激素治疗的试剂。这种激素调控试剂的实施例有它莫西芬、艾多昔芬、氟维司群、屈洛昔芬、托瑞米芬、雷洛昔芬、己烯雌酚、乙炔基雌二醇/乙炔雌二醇、抗雄激素(譬如氟他胺)、孕激素(譬如羟基孕酮己酸盐、甲孕酮/安宫黄体酮、甲地孕酮醋酸盐/美可治)、肾上腺皮质激素(譬如氢化可的松、强的松)、黄体化激素释放激素(及其类似物和其它LHRH拮抗剂，例如布舍瑞林和戈舍瑞林)、芳香酶抑制剂(例如瑞宁得/瑞宁得、奥美定/cytraden、依西美坦)、激素抑制剂(例如奥曲肽/-善得定)和类似试剂。In one technical solution, the therapeutic agent used in combination with the CD38BPs of the present invention to treat the above-mentioned diseases is a hormone-modulating agent, such as an agent for anti-androgen and anti-estrogen therapy. Examples of such hormone modulating agents are tamoxifen, edoxifene, fulvestrant, droloxifene, toremifene, raloxifene, diethylstilbestrol, ethinyl estradiol/ethinyl estradiol Alcohols, antiandrogens (eg, flutamide), progestins (eg, hydroxyprogesterone caproate, medroxyprogesterone/uterine progesterone, megestrol acetate/mecox), adrenal corticosteroids (eg, hydrocortisone, prednisone), luteinizing hormone-releasing hormone (and its analogs and other LHRH antagonists such as buserelin and goserelin), aromatase inhibitors (such as Arimidex/Arimidex octreotide/cytraden, exemestane), hormone inhibitors (eg, octreotide/-sandodine), and similar agents.

在一个技术方案中，与本发明的CD38BPs联合使用以治疗上述疾病的治疗性试剂是抗无反应性化疗剂(例如耐受肿瘤和癌抗原的小分子化合物、蛋白、糖蛋白或抗体)。这种化合物的实施例是可阻断CTLA-4活性的分子，例如MDX-010(Phan et al.，PNAS USA 100，8372(2003))。In one technical solution, the therapeutic agent used in combination with CD38BPs of the present invention to treat the above-mentioned diseases is an anti-anergy chemotherapeutic agent (such as a small molecule compound, protein, glycoprotein or antibody resistant to tumor and cancer antigens). Examples of such compounds are molecules that block CTLA-4 activity, such as MDX-010 (Phan et al., PNAS USA 100, 8372 (2003)).

在一个技术方案中，与本发明的CD38BPs联合使用以治疗上述疾病的治疗性试剂是含肿瘤抑制子基因核酸或诸如编码人重组野生型p53/SCH58500的复制缺陷型腺病毒这样的载体等；针对癌基因的反义核酸或去调控基因；或针对突变或去调控基因的siRNA。肿瘤抑制子靶的实施例包括例如BRCA1、RB1、BRCA2、DPC4(Smad4)、MSH2、MLH1和DCC。In a technical scheme, the therapeutic agent used in combination with CD38BPs of the present invention to treat the above-mentioned diseases is a nucleic acid containing a tumor suppressor gene or a vector such as a replication-deficient adenovirus encoding human recombinant wild-type p53/SCH58500; Antisense to oncogenes or deregulated genes; or siRNA to mutated or deregulated genes. Examples of tumor suppressor sub-targets include, eg, BRCA1, RB1, BRCA2, DPC4 (Smad4), MSH2, MLH1, and DCC.

在一个技术方案中，与本发明的CD38BPs联合使用以治疗上述疾病的治疗性试剂是抗癌核酸，例如genasense(augmerosen/G3139)、LY900003(ISIS 3521)、ISIS 2503、OGX-011(ISIS 112989)、LE- AON/LEraf-AON(脂质体胶囊化的c-raf反义寡核苷酸/ISIS-5132)、MG98和其它针对PKCα、丛生蛋白、IGFBPs、白蛋白激酶A、细胞周期蛋白D1或Bcl-2h的反义核酸。In a technical solution, the therapeutic agent used in combination with CD38BPs of the present invention to treat the above-mentioned diseases is an anticancer nucleic acid, such as genasense (augmerosen/G3139), LY900003 (ISIS 3521), ISIS 2503, OGX-011 (ISIS 112989) , LE-AON/LEraf-AON (liposome-encapsulated c-raf antisense oligonucleotide/ISIS-5132), MG98 and others targeting PKCα, clusterin, IGFBPs, albumin kinase A, cyclin D1 Or the antisense nucleic acid of Bcl-2h.

在一个技术方案中，与本发明的CD38BPs联合使用以治疗上述疾病的治疗性试剂是抗癌抑制性RNA分子(参见例如Lin et al.，CurrCancer Drug Targets.1(3)，241-7(2001)、Erratum in：Curr Cancer DrugTargets.3(3)，237(2003)、Lima et al.，CancerGene Ther.11(5)，309-16(2004)、Grzmil et al.，lnt J Oncol.4(1)，97-105(2004)、Collis et al.，lnt J Radiat Oncol Biol Phys.57(2 Suppl)，S144(2003)、Yang etal.，Oncogene.22(36)，5694-701(2003)和Zhang et al.，BiochemBiophys ResCommun.303(4)，1169-78(2003))。In one technical solution, the therapeutic agent used in combination with CD38BPs of the present invention to treat the above-mentioned diseases is an anti-cancer inhibitory RNA molecule (see for example Lin et al., CurrCancer Drug Targets.1 (3), 241-7 (2001 ), Erratum in: Curr Cancer Drug Targets.3(3), 237(2003), Lima et al., CancerGene Ther.11(5), 309-16(2004), Grzmil et al., lnt J Oncol.4( 1), 97-105(2004), Collis et al., lnt J Radiat Oncol Biol Phys.57(2 Suppl), S144(2003), Yang et al., Oncogene.22(36), 5694-701(2003) and Zhang et al., Biochem Biophys Res Commun. 303(4), 1169-78 (2003)).

本发明的组分和联合给药也包括给药核酸疫苗，例如编码癌抗原/肿瘤相关抗原的裸露DNA疫苗(参见例如，US 5,589,466、US5,593,972、US 5,703,057、US 5,879,687、US6,235,523和US 6,387,888)。在一个技术方案中，联合给药方法和/或联合组分包括自体疫苗组分。在一个技术方案中，联合给药方法和/或联合组分包括全细胞疫苗或细胞因子表达细胞(例如重组IL-2表达成纤维细胞、重组细胞因子表达枝状细胞等)(参见例如，Kowalczyk et al.，Acta Biochim Pol.50(3)，613-24(2003)、Reilly et al.，MethodsMoI Med.69，233-57(2002)和Tirapuet al.，Curr Gene Ther.2(1)，79-89(2002)。这种自体细胞技术用于联合方法的另一个实施例是MyVax个体化免疫治疗方法(以前称为GTOP-99)(Genitope Corporation-Redwood City，CA，USA)。The compositions and combinations of the present invention also include the administration of nucleic acid vaccines, such as naked DNA vaccines encoding cancer antigens/tumor-associated antigens (see, for example, US 5,589,466, US 5,593,972, US 5,703,057, US 5,879,687, US 6,235,523 and US 6,387,888). In one technical solution, the combination administration method and/or combination components include autologous vaccine components. In one technical solution, the combined administration method and/or combined components include whole cell vaccines or cytokine expressing cells (such as recombinant IL-2 expressing fibroblasts, recombinant cytokine expressing dendritic cells, etc.) (see for example, Kowalczyk et al., Acta Biochim Pol.50(3), 613-24(2003), Reilly et al., MethodsMoI Med.69, 233-57(2002) and Tirapu et al., Curr Gene Ther.2(1), 79-89 (2002).Another example of this autologous cell technology being used in a combined approach is MyVax Personalized immunotherapy approach (formerly known as GTOP-99) (Genitop Corporation-Redwood City, CA, USA).

在一个技术方案中，本发明提供联合组分和联合给药方法，其中CD38BP与病毒、病毒蛋白等联合给药或共同给药。例如，通常仅具有一轮或几轮体内复制并定位于肿瘤细胞的复制缺陷型病毒可用作这种组分和方法的成分。这种病毒试剂包括或与编码免疫刺激剂，例如GM-CSF和/或IL-2的核酸相关。天然溶瘤和重组溶瘤病毒(例如HSV-1病毒、呼肠孤病毒、复制缺陷型和复制敏感型腺病毒等)可用作这种方法和组分的成分。因此，在一个技术方案中，本发明提供联合组分和联合给药方法，其中CD38BP与溶瘤病毒联合给药或共同给药。这种病毒的实施例包括溶瘤腺病毒和疱疹病毒，它们可以是修饰或不进行修饰的病毒(参见例如，Shah et al.，J Neurooncol.65(3)，203-26(2003)、 Stiles et al.，Surgery.134(2)，357-64(2003)、Sunarmura et al.，Pancreas.28(3)，326-9(2004)、Teshigahara et al.，J Surg Oncol.85(1)，42-7(2004)、Varghese et al.，Cancer GeneTher.9(12)，967-78(2002)、Wildner et al.，Cancer Res.59(2)，410-3(1999)、Yamanaka，lnt J Oncol.24(4)，919-23(2004)和Zwiebel et al.，Semin Oncol.28(4)，336-43(2001)。In one technical solution, the present invention provides combined components and a combined administration method, wherein CD38BP is administered or co-administered with viruses, viral proteins and the like. For example, replication-defective viruses that normally have only one or a few rounds of replication in vivo and localize to tumor cells can be used as components of such components and methods. Such viral agents include or are associated with nucleic acids encoding immunostimulatory agents, such as GM-CSF and/or IL-2. Natural and recombinant oncolytic viruses (eg, HSV-1 virus, reovirus, replication-deficient and replication-sensitive adenoviruses, etc.) can be used as components of such methods and components. Therefore, in a technical solution, the present invention provides combined components and combined administration methods, wherein CD38BP is administered or co-administered with oncolytic virus. Examples of such viruses include oncolytic adenoviruses and herpes viruses, which may be modified or unmodified viruses (see, e.g., Shah et al., J Neurooncol. 65(3), 203-26 (2003), Stiles et al., Surgery.134(2), 357-64(2003), Sunarmura et al., Pancreas.28(3), 326-9(2004), Teshigahara et al., J Surg Oncol.85(1) , 42-7(2004), Varghese et al., Cancer GeneTher.9(12), 967-78(2002), Wildner et al., Cancer Res.59(2), 410-3(1999), Yamanaka, lnt J Oncol. 24(4), 919-23(2004) and Zwiebel et al., Semin Oncol. 28(4), 336-43(2001).

本发明的联合组分和联合给药方法也包括“全细胞”和“过继”免疫治疗方法。例如，这种方法包括免疫系统细胞(例如肿瘤浸润淋巴细胞(TILs)、例如CD4+和/或CD8+T细胞(例如具有肿瘤特异抗原和/或基因增强的T细胞)的灌液或再次灌液、抗体表达B细胞和其它抗体产生/呈递细胞、枝状细胞(例如，抗细胞因子病毒重组枝状细胞、用诸如GM-CSF和/或Flt3-L这样的DC扩大试剂培养的枝状细胞、和/或肿瘤相关抗原负载枝状细胞)、抗肿瘤NK细胞、所谓的杂合细胞或其组合。细胞裂解物也用于这种方法和组分中。可用于这方面的临床实验中的细胞型“疫苗”包括Canvaxin^TM、APC-8015(Dendreon)、HSPPC-96(Antigenics)和Melacine细胞裂解物。与佐剂，例如明矾混合的从癌细胞脱落的抗原及其混合物(参见例如Bystryn et al.，Clinical CancerResearch Vol.7，1882-1887，July 2001)选择性地也是这种方法和联合组分这的成分。The combination components and methods of combination administration of the invention also include "whole cell" and "adoptive" immunotherapy methods. For example, such methods include perfusion or reperfusion of immune system cells such as tumor infiltrating lymphocytes (TILs), such as CD4+ and/or CD8+ T cells such as T cells with tumor-specific antigens and/or genetic enhancement , antibody expressing B cells and other antibody producing/presenting cells, dendritic cells (e.g., anti-cytokine virus recombinant dendritic cells, dendritic cells cultured with DC expansion reagents such as GM-CSF and/or Flt3-L, and/or tumor-associated antigen-loaded dendritic cells), anti-tumor NK cells, so-called hybrid cells, or combinations thereof. Cell lysates are also used in this method and components. Cells that can be used in clinical trials in this regard Type "vaccine" includes Canvaxin ^TM , APC-8015 (Dendreon), HSPPC-96 (Antigenics) and Melacine cell lysate. Antigens shed from cancer cells and mixtures thereof mixed with adjuvants, such as alum (see for example Bystryn et al., Clinical Cancer Research Vol.7, 1882-1887, July 2001) are optionally also this method and combination components. ingredients.

在一个技术方案中，本发明的CD38BP可与内部接种疫苗的方法联合递送到患者中。内部接种疫苗是指在患者中，诱导的肿瘤细胞或癌细胞死亡，例如药物诱导或辐射诱导的肿瘤细胞的细胞死亡，典型地引发针对以下的免疫应答(i)作为整体的肿瘤细胞，或(ii)包括以下的肿瘤细胞部分(a)分泌蛋白、蛋白或其它产物，(b)膜相关蛋白或糖蛋白或其它膜相关或插入膜中的组分，和/或(c)细胞内蛋白或其它细胞内成分。内部接种疫苗诱导的免疫应答可是由体液引起的(即抗体-补体介导)或细胞介导的(例如，识别内部被杀死的肿瘤细胞或其部分的内源细胞毒T淋巴细胞的发育和/或增加)。除了放射性治疗外，可用于诱导所述肿瘤细胞死亡和内部免疫化的药物和试剂的非限制性实施例是传统的化疗试剂、细胞循环抑制剂、抗血管生成药物、单克隆抗体、凋亡诱导试剂和信号转导抑制剂。与本发明的CD38BPs联合使用作为治疗性试剂相关的治疗上述疾病的其它抗癌试剂的实施例是分化诱导试 剂、维A酸和维A酸类似物(例如全反式维A酸、13-顺式维A酸及类似试剂)、维生素D类似物(例如西奥骨化醇及类似试剂)、ErbB3、ErbB4、IGF-IR抑制剂、胰岛素受体、PDGFRα、PDGFRβ、Flk2、Flt4、FGFR1、FGFR2、FGFR3、FGFR4、TRKA、TRKC、c-met、Ron、Sea、Tie、Tie2、Eph、Ret、Ros、AIk、LTK、PTK7及类似试剂。In one embodiment, the CD38BP of the present invention can be delivered to a patient in combination with internal vaccination. Internal vaccination means that in a patient, induced tumor cells or cancer cell death, such as drug-induced or radiation-induced cell death of tumor cells, typically elicits an immune response against (i) the tumor cells as a whole, or ( ii) tumor cell fractions comprising (a) secreted proteins, proteins or other products, (b) membrane-associated proteins or glycoproteins or other membrane-associated or membrane-inserted components, and/or (c) intracellular proteins or other intracellular components. The immune response induced by internal vaccination can be humoral (i.e., antibody-complement mediated) or cell-mediated (e.g., the development and / or increase). In addition to radiotherapy, non-limiting examples of drugs and agents that can be used to induce death and internal immunization of the tumor cells are traditional chemotherapeutic agents, cell cycle inhibitors, anti-angiogenic drugs, monoclonal antibodies, apoptosis-inducing Reagents and Signal Transduction Inhibitors. Examples of other anticancer agents for the treatment of the above-mentioned diseases that are used in combination with CD38BPs of the present invention as therapeutic agents are differentiation-inducing agents, retinoic acid and retinoic acid analogs (such as all-trans retinoic acid, 13-cis retinoic acid and similar agents), vitamin D analogs (such as ceocalcitol and similar agents), ErbB3, ErbB4, IGF-IR inhibitors, insulin receptors, PDGFRα, PDGFRβ, Flk2, Flt4, FGFR1, FGFR2 , FGFR3, FGFR4, TRKA, TRKC, c-met, Ron, Sea, Tie, Tie2, Eph, Ret, Ros, AIk, LTK, PTK7 and similar reagents.

与本发明的CD38BPs联合使用作为治疗性试剂相关的治疗上述疾病的其它抗癌试剂的实施例有组织蛋白酶B、组织蛋白酶D脱氢酶活性的调控子、谷光甘肽-S-转移酶(例如谷氨酰半胱氨酸合成酶和乳酸脱氢酶)及类似试剂。Examples of other anticancer agents for the treatment of the above-mentioned diseases that are used in combination with CD38BPs of the present invention as therapeutic agents include regulators of cathepsin B, cathepsin D dehydrogenase activity, glutathione-S-transferase (such as glutamylcysteine synthetase and lactate dehydrogenase) and similar reagents.

与本发明的CD38BPs联合使用作为治疗性试剂相关的治疗上述疾病的其它抗癌试剂的实施例有雌莫司汀和表柔比星。Examples of other anticancer agents for the treatment of the above-mentioned diseases that are used in combination with CD38BPs of the present invention as therapeutic agents are estramustine and epirubicin.

与本发明的CD38BPs联合使用作为治疗性试剂相关的治疗上述疾病的其它抗癌试剂的实施例有HSP90抑制剂类似的1 7-丙烯基氨基格尔德霉素、针对诸如PSA1 CA125、KSA等肿瘤抗原的抗体、整联蛋白类似的整联蛋白β1、VCAM抑制剂及类似试剂。Examples of other anti-cancer agents used in combination with CD38BPs of the present invention as therapeutic agents for the treatment of the above-mentioned diseases include 1 7-propenylaminogeldanamycin, which is similar to HSP90 inhibitors, for tumors such as PSA1 CA125, KSA, etc. Antibodies to antigens, integrin-like integrin beta 1 , VCAM inhibitors, and similar agents.

与本发明的CD38BPs联合使用作为治疗性试剂相关的治疗上述疾病的其它抗癌试剂的实施例有神经钙蛋白抑制剂(例如伐司朴达、PSC833和其它MDR-1或p-糖蛋白抑制剂)，TOR抑制剂(例如西罗莫司、依维莫司和雷帕霉素)。及“淋巴细胞归巢”机制的抑制剂(例如FTY720)及影响细胞信号转导的抑制剂，例如黏附分子抑制剂(例如LFA等)。Examples of other anticancer agents for the treatment of the above-mentioned diseases that are used in combination with CD38BPs of the present invention as therapeutic agents include calcineurin inhibitors (such as valspodar, PSC833 and other MDR-1 or p-glycoprotein inhibitors ), TOR inhibitors (such as sirolimus, everolimus, and rapamycin). And inhibitors of "lymphocyte homing" mechanism (such as FTY720) and inhibitors affecting cell signal transduction, such as adhesion molecule inhibitors (such as LFA, etc.).

在一个技术方案中，本发明提供在患者中治疗由表达CD38的细胞参与的疾病的方法，该方法包括给药所需患者治疗有效剂量的本发明的CD38BP和放射性治疗。In one embodiment, the present invention provides a method of treating a disease involving CD38-expressing cells in a patient, the method comprising administering a therapeutically effective dose of a CD38BP of the invention and radiotherapy to the patient in need.

在一个技术方案中，本发明提供在患者中治疗多发性骨髓瘤的方法，该方法包括给药所需患者治疗有效剂量的本发明的CD38BP和放射性治疗。In one embodiment, the present invention provides a method of treating multiple myeloma in a patient, the method comprising administering a therapeutically effective dose of CD38BP of the present invention and radiation therapy to the patient in need.

在一个技术方案中，本发明提供本发明的CD38BP在制备与放射性治疗联合使用以治疗多发性骨髓瘤的药物组合物中的用途。In one technical solution, the present invention provides the use of the CD38BP of the present invention in the preparation of a pharmaceutical composition used in combination with radiotherapy to treat multiple myeloma.

放射性治疗可包括对患者提供的放射性药剂的辐射或相关的给药。辐射的来源对被治疗的患者来说可以外部或内部的(例如，辐射治疗可以外部光束治疗(EBRT)、短程治疗(BT)或针对骨骼的放射性 治疗形式)。用于这种方法的放射性元素包括，例如镭、铯-137、铱-192、镅-241、金-198、钴-57、铜-67、锝-99、碘-123、碘-131和铟-111。Radiation therapy may include radiation or related administration of radiopharmaceuticals provided to the patient. The source of radiation can be external or internal to the patient being treated (for example, radiation therapy can be in the form of external beam therapy (EBRT), brachytherapy (BT), or bone-targeted radiation therapy). Radioactive elements used in this method include, for example, radium, cesium-137, iridium-192, americium-241, gold-198, cobalt-57, copper-67, technetium-99, iodine-123, iodine-131, and indium -111.

在一个技术方案中，本发明提供在患者中治疗由表达CD38的细胞参与的疾病的方法，该方法包括给药所需患者治疗有效剂量的本发明的CD38BP和自体固有的外周干细胞或骨髓移植。In one technical solution, the present invention provides a method for treating a disease involving CD38-expressing cells in a patient, the method comprising administering a therapeutically effective dose of CD38BP of the present invention and autologous peripheral stem cells or bone marrow transplantation to the patient in need.

在一个技术方案中，本发明提供在患者中治疗多发性骨髓瘤的方法，该方法包括给药所需患者治疗有效剂量的本发明的CD38BP和自体固有的外周干细胞或骨髓移植。In one technical solution, the present invention provides a method for treating multiple myeloma in a patient, the method comprising administering a therapeutically effective dose of CD38BP of the present invention and autologous peripheral stem cells or bone marrow transplantation to the patient in need.

在一个技术方案中，本发明提供本发明的CD38BP在制备与自体固有的外周干细胞或骨髓移植联合使用以治疗多发性骨髓瘤的药物组合物中的用途。In one technical solution, the present invention provides the use of the CD38BP of the present invention in the preparation of a pharmaceutical composition used in combination with autologous peripheral stem cells or bone marrow transplantation to treat multiple myeloma.

在一个技术方案中，本发明提供在患者中治疗由表达CD38的细胞参与的疾病的方法，该方法包括给药所需患者治疗有效剂量的本发明的CD38BP和整形外科干涉。In one embodiment, the present invention provides a method of treating a disease involving CD38-expressing cells in a patient, the method comprising administering to the patient in need thereof a therapeutically effective dose of a CD38BP of the invention and orthopedic intervention.

在一个技术方案中，本发明提供本发明的CD38BP在制备与自体固有的外周干细胞或骨髓移植联合使用以治疗多发性骨髓瘤的药物组合物中的用途。In one technical solution, the present invention provides the use of the CD38BP of the present invention in the preparation of a pharmaceutical composition used in combination with autologous peripheral stem cells or bone marrow transplantation to treat multiple myeloma.

整形外科干涉可用于治疗由表达CD38的细胞参与的疾病，例如多发性骨髓瘤，以协助控制疼痛或维持功能或活动性。这种干涉包括康复治疗、骨骼夹板以防止骨折、或外科手术(次要或主要)以修复骨折。Orthopedic interventions can be used to treat diseases involving CD38-expressing cells, such as multiple myeloma, to help manage pain or maintain function or mobility. Such interventions include rehabilitation, bone splinting to prevent fractures, or surgery (minor or major) to repair fractures.

在一个技术方案中，本发明的CD38BP可与一种或多种促进CD38BP或组合物接近肿瘤内部的递送联合给药。例如，这种方法可与能松弛肿瘤的松弛肽联合递送(参见例如US6,719,977)。在一个技术方案中，本发明的CD38BP可与细胞穿透肽(CPP)结合。细胞穿透肽和相关的肽(例如构建的细胞穿透抗体)在例如Zhao et al.，J ImmunolMethods.254(1-2)，137-45(2001)、Hong et al.，Cancer Res.60(23)，6551-6(2000).Lindgren et al.，Biochem J.377(Pt1)、69-76(2004)、Buerger et al.，J Cancer Res Clin Oncol.129(12)，669-75(2003)、Poogaet al.，FASEB J.12(1)，67-77(1998)和Tseng et al.，MoIPharmacol.62(4)，864-72(2002)中描述。In one technical solution, the CD38BP of the present invention can be administered in combination with one or more methods that promote the delivery of CD38BP or the composition close to the interior of the tumor. For example, this approach can be delivered in conjunction with a relaxin peptide that relaxes tumors (see eg US 6,719,977). In one technical solution, the CD38BP of the present invention can be combined with a cell penetrating peptide (CPP). Cell penetrating peptides and related peptides (e.g. constructed cell penetrating antibodies) are described in, for example, Zhao et al., J Immunol Methods. 254(1-2), 137-45 (2001), Hong et al., Cancer Res.60 (23), 6551-6(2000). Lindgren et al., Biochem J. 377(Pt1), 69-76(2004), Buerger et al., J Cancer Res Clin Oncol. 129(12), 669-75 (2003), Pooga et al., FASEB J.12(1), 67-77 (1998) and Tseng et al., MoI Pharmacol. 62(4), 864-72 (2002).

在一个技术方案中，本发明提供在患者中治疗由表达CD38的细胞参与的疾病的方法，该方法包括给药所需患者治疗有效剂量的本发明的 CD38BP和至少一种抗炎症试剂。In one embodiment, the invention provides a method of treating a disease involving CD38-expressing cells in a patient, the method comprising administering to the patient in need thereof a therapeutically effective amount of a CD38BP of the invention and at least one anti-inflammatory agent.

在一个技术方案中，本发明提供在患者中治疗多发性骨髓瘤的方法，该方法包括给药所需患者治疗有效剂量的本发明的CD38BP和至少一种抗炎症试剂。In one embodiment, the invention provides a method of treating multiple myeloma in a patient, the method comprising administering to the patient in need thereof a therapeutically effective amount of a CD38BP of the invention and at least one anti-inflammatory agent.

在一个技术方案中，本发明提供本发明的CD38BP在制备与至少一种抗炎症试剂联合使用以治疗多发性骨髓瘤的药物组合物中的用途。In one technical solution, the present invention provides the use of the CD38BP of the present invention in the preparation of a pharmaceutical composition used in combination with at least one anti-inflammatory agent to treat multiple myeloma.

在一个技术方案中，这种抗炎症试剂选自类固醇药物和NSAID(非类固醇抗炎症药物)。In a technical solution, this anti-inflammatory agent is selected from steroid drugs and NSAIDs (non-steroidal anti-inflammatory drugs).

在一个技术方案中，这种抗炎症试剂选自用于治疗炎症性疾病的阿司匹林和其它水杨酸、Cox-2抑制物(譬如罗非考昔和塞来考昔)、NSAIDs(譬如布洛芬、酮洛芬、萘普生、枢力达、双氯芬酸、吡罗昔康、酮洛芬、二氟尼柳、萘丁美酮、依托度酸、奥沙普秦和消炎痛)、抗IL6R抗体、抗IL8抗体、抗体IL15抗体、抗IL15R抗体、抗CD4抗体、抗CD11a抗体(例如依法利珠单抗)、抗-α-4/β-1整联蛋白(VLA4)抗体(例如那他珠单抗)、CTLA4-Ig、氢化泼尼松、强的松、诸如甲氨蝶呤这样的病症缓解性抗风湿药(DMARDs)、羟化氯喹、柳氮磺胺吡啶、嘧啶合成抑制物(譬如来氟米特)、IL-1受体阻断试剂(譬如阿那白滞素)、TNF-α阻断试剂(譬如依那西普、英夫利昔单抗和阿达木单抗)及类似试剂。In one embodiment, the anti-inflammatory agent is selected from aspirin and other salicylates used in the treatment of inflammatory diseases, Cox-2 inhibitors (such as rofecoxib and celecoxib), NSAIDs (such as buprofen, fen, ketoprofen, naproxen, citrita, diclofenac, piroxicam, ketoprofen, diflunisal, nabumetone, etodolac, oxaprozin and indomethacin), anti-IL6R antibody, Anti-IL8 antibody, anti-IL15 antibody, anti-IL15R antibody, anti-CD4 antibody, anti-CD11a antibody (such as efalizumab), anti-α-4/β-1 integrin (VLA4) antibody (such as natalizumab anti-), CTLA4-Ig, prednisone, prednisone, disease-modifying antirheumatic drugs (DMARDs) such as methotrexate, hydroxychloroquine, sulfasalazine, pyrimidine synthesis inhibitors (such as leflu Miter), IL-1 receptor blocking agents (such as anakinra), TNF-α blocking agents (such as etanercept, infliximab, and adalimumab), and similar agents.

在一个技术方案中，本发明提供在患者中治疗由表达CD38的细胞参与的疾病的方法，该方法包括给药所需患者治疗有效剂量的本发明的CD38BP和至少一种免疫抑制剂和/或免疫调节剂。In one technical solution, the present invention provides a method for treating a disease involving CD38-expressing cells in a patient, the method comprising administering a therapeutically effective dose of CD38BP of the present invention and at least one immunosuppressant and/or Immunomodulators.

在一个技术方案中，本发明提供在患者中治疗多发性骨髓瘤的方法，该方法包括给药所需患者治疗有效剂量的本发明的CD38BP和至少一种免疫抑制剂和/或免疫调节剂。In one technical aspect, the present invention provides a method of treating multiple myeloma in a patient, the method comprising administering a therapeutically effective dose of CD38BP of the present invention and at least one immunosuppressant and/or immunomodulator to the patient in need.

在一个技术方案中，本发明提供本发明的CD38BP在制备与至少一种免疫抑制剂和/或免疫调节剂联合使用以治疗多发性骨髓瘤的药物组合物中的用途。In one technical solution, the present invention provides the use of the CD38BP of the present invention in the preparation of a pharmaceutical composition used in combination with at least one immunosuppressant and/or immunomodulator to treat multiple myeloma.

在一个技术方案中，这种免疫抑制剂和/或免疫调节剂选自环孢霉素、咪唑硫嘌呤、霉酚酸、麦考酚酸莫酯、譬如强的松这样的皮脂类固醇、甲氨蝶呤、金盐、柳氮磺胺吡啶、抗疟药、布喹那、来氟米特、咪唑立宾、脱氧精胍菌素、6-巯基嘌呤、环磷酰胺、雷帕霉素、他克莫司 (FK-506)、OKT3、抗胸腺细胞球蛋白、胸腺五肽、胸腺素-α及类似试剂。In a technical solution, this immunosuppressant and/or immunomodulator is selected from cyclosporine, azathioprine, mycophenolic acid, mycophenolate mofetil, corticosteroids such as prednisone, methylamine Pterin, gold salt, sulfasalazine, antimalarials, buquinar, leflunomide, mizoribine, deoxyspergualin, 6-mercaptopurine, cyclophosphamide, rapamycin, tagram Limus (FK-506), OKT3, antithymocyte globulin, thymopentin, thymosin-α, and similar agents.

在一个技术方案中，这种免疫抑制剂和/或免疫调节剂选自免疫抑制性抗体，例如与IL-2受体的p75结合的抗体，或与例如MHC、CD2、CD3、CD4、CD7、CD28、B7、CD40、CD45、IFNγ、TNF-α、IL-4、IL-5、IL-6R、IL-6；IGF、IGFR1、IL-7、IL-8、IL-10、CD11a或CD58结合的抗体，或与其配体结合的抗体。In one technical solution, this immunosuppressant and/or immunomodulator is selected from immunosuppressive antibodies, such as antibodies that bind to p75 of the IL-2 receptor, or antibodies that bind to, for example, MHC, CD2, CD3, CD4, CD7, CD28, B7, CD40, CD45, IFNγ, TNF-α, IL-4, IL-5, IL-6R, IL-6; IGF, IGFR1, IL-7, IL-8, IL-10, CD11a, or CD58 binding antibody, or an antibody that binds to its ligand.

在一个技术方案中，这种免疫抑制剂和/或免疫调节剂选自可溶性IL-15R、IL-10、B7分子(B7-1、B7-2、其变异体及其片段)、ICOS和OX40、T细胞负调控因子抑制剂(例如抗CTLA4抗体)及类似试剂。In one technical solution, this immunosuppressant and/or immunomodulator is selected from soluble IL-15R, IL-10, B7 molecules (B7-1, B7-2, variants and fragments thereof), ICOS and OX40 , T cell negative regulator inhibitors (such as anti-CTLA4 antibodies) and similar reagents.

在一个技术方案中，本发明的CD38BP可与两种或多种免疫抑制剂和/或免疫调节剂联合给药，例如与强的松和环孢霉素；强的松、环孢霉素和咪唑硫嘌呤；或强的松、环孢霉素和麦考酚酸莫酯联合给药。In a technical scheme, CD38BP of the present invention can be administered in combination with two or more immunosuppressants and/or immunomodulators, for example with prednisone and cyclosporine; prednisone, cyclosporine and Azathioprine; or a combination of prednisone, cyclosporine, and mycophenolate mofetil.

在一个技术方案中，本发明提供在患者中治疗由表达CD38的细胞参与的疾病的方法，该方法包括给药所需患者治疗有效剂量的本发明的CD38BP和抗C3b(i)抗体。In one embodiment, the present invention provides a method of treating a disease involving CD38-expressing cells in a patient, the method comprising administering to the patient in need thereof a therapeutically effective dose of a CD38BP of the invention and an anti-C3b(i) antibody.

在一个技术方案中，本发明提供在患者中治疗多发性骨髓瘤的方法，该方法包括给药所需患者治疗有效剂量的本发明的CD38BP和抗C3b(i)抗体。In one embodiment, the present invention provides a method of treating multiple myeloma in a patient, the method comprising administering to the patient in need thereof a therapeutically effective amount of CD38BP and an anti-C3b(i) antibody of the present invention.

在一个技术方案中，本发明提供本发明的CD38BP在制备与抗C3b(i)抗体联合使用以治疗多发性骨髓瘤的药物组合物中的用途。In one technical solution, the present invention provides the use of the CD38BP of the present invention in the preparation of a pharmaceutical composition used in combination with an anti-C3b(i) antibody to treat multiple myeloma.

在一个技术方案中，与本发明的CD38BPs联合使用以治疗上述疾病的治疗试剂选自组蛋白去乙酰基转移酶抑制剂(例如苯基丁酸)和/或DNA修复试剂(例如DNA修复酶和相关组分，譬如dimericine)。In one technical solution, the therapeutic agent used in combination with CD38BPs of the present invention to treat the above-mentioned diseases is selected from histone deacetyltransferase inhibitors (such as phenylbutyric acid) and/or DNA repair agents (such as DNA repair enzymes and related components, such as dimericine).

包括给药治疗有效剂量的本发明的CD38BP以治疗上述疾病的方法也可包括抗癌定向光敏疗法(例如抗癌激光治疗，它选择性地可使用光敏试剂，参见例如，Zhang et al.，J Control Release.93(2)，141-50(2003))、抗癌声波和冲击波治疗(例如参见，Kambe etal.，Hum Cell.10(1)，87-94(1997))，和/或抗癌营养辅助食品治疗(参见例如，Roudebushet al.，Vet Clin North Am Small Anim Pract.34(U 249-69，viii(2004)和Rafi，Nutrition.20(1)，78-82(2004)。同样，本发明的CD38BP可用作制备治疗上述疾病的药物组合物，与抗癌定向的光力学 治疗联合给药(例如，抗癌激光治疗，它选择性地可使用光敏试剂，抗癌声波和冲击波治疗，和/或抗癌营养辅助食品治疗)。The method comprising administering a therapeutically effective dose of the CD38BP of the present invention to treat the above-mentioned diseases may also include anti-cancer directed photosensitization therapy (such as anti-cancer laser therapy, which optionally uses photosensitizing agents, see, e.g., Zhang et al., J Control Release.93(2), 141-50(2003)), anticancer sonic and shock wave therapy (see, for example, Kambe et al., Hum Cell.10(1), 87-94(1997)), and/or anticancer Cancer Nutritional Supplementary Food Therapy (see for example, Roudebush et al., Vet Clin North Am Small Anim Pract. 34 (U 249-69, viii (2004) and Rafi, Nutrition. 20(1), 78-82 (2004). Likewise , the CD38BP of the present invention can be used as a pharmaceutical composition for the preparation of the above-mentioned diseases, and it can be administered in combination with anti-cancer directed photodynamic therapy (for example, anti-cancer laser therapy, which can selectively use photosensitizers, anti-cancer sound waves and shock waves) treatment, and/or anticancer nutritional supplements for food therapy).

如上所述，本发明的药物组合物可联合给药进行治疗，即，作为单独的药物组合物或与一种或多种其它上述治疗性试剂一起配制的本发明的化合物与一种或多种与被治疗疾病或症状相关的试剂联合使用。这种联合治疗需要更低剂量的本发明的化合物和/或共给药试剂，从而避免与各种单一治疗相关的可能毒性或并发症。As noted above, the pharmaceutical compositions of the present invention may be administered in combination therapy, i.e., a compound of the present invention and one or more Used in combination with agents related to the disease or condition being treated. Such combination therapy requires lower doses of the compounds of the invention and/or co-administered agents, thereby avoiding possible toxicity or complications associated with various monotherapies.

在一个技术方案中，本发明提供与免疫调节物偶联的CD38BP，譬如免疫调节细胞因子、干细胞生长因子、淋巴毒素(例如TNF，譬如TNFα)，或造血因子。这种可用作连接物的分子的实施例包括IL-1、IL-2、IL-3、IL-6、IL-10、IL-12、IL-18和IL-21、集落刺激因子(例如粒细胞集落刺激因子(G-CSF)和粒细胞-巨噬细胞集落刺激因子(GM-CSF))、干扰素(例如IFNα、IFNβ和IFNy)、命名为“S1因子”的干细胞生长因子、促红细胞生成素和促血小板生成素、其活性片段、其衍生物、其变异体或任何其组合。In one embodiment, the present invention provides CD38BP conjugated to an immune modulator, such as an immunomodulatory cytokine, stem cell growth factor, lymphotoxin (eg TNF, such as TNFα), or a hematopoietic factor. Examples of such molecules that can be used as linkers include IL-1, IL-2, IL-3, IL-6, IL-10, IL-12, IL-18 and IL-21, colony stimulating factors (e.g. Granulocyte-colony-stimulating factor (G-CSF) and granulocyte-macrophage-colony-stimulating factor (GM-CSF)), interferons (such as IFNα, IFNβ, and IFNy), stem cell growth factors named "S1 factors", pro- Erythropoietin and thrombopoietin, active fragments thereof, derivatives thereof, variants thereof or any combination thereof.

在一个技术方案中，本发明的CD38BPs可通过检测其膜表面的CD38水平或含CD38的细胞水平，从而在体内或体外诊断其中激活的表达CD38的细胞在发病机理中起重要作用的疾病。例如，可通过将待测样品，选择性地包括对照样品与CD38BP在抗体和CD38间可形成复合物的条件下接触。然后检测复合物的形成(例如使用ELISA)。当对照样品与检测样品一起使用时，在两种样品中检测复合物，两种样品复合物形成的任何统计学上显著的差异即表明在检测样品中存在CD38。In a technical scheme, the CD38BPs of the present invention can detect the CD38 level on its membrane surface or the level of CD38-containing cells, thereby in vivo or in vitro diagnosis of diseases in which activated CD38-expressing cells play an important role in pathogenesis. For example, a test sample, optionally including a control sample, can be contacted with CD38BP under conditions under which a complex can be formed between the antibody and CD38. Complex formation is then detected (eg using ELISA). When a control sample is used with a test sample, the complex is detected in both samples and any statistically significant difference in complex formation between the two samples is indicative of the presence of CD38 in the test sample.

更具体地，本发明提供鉴定和诊断入侵细胞和组织及其它本发明的CD38BPs定位的细胞，从而监测治疗进展、治疗后的状态、发展癌症的危险性、癌症进程等。在一个这种诊断检测的实施例中，本发明提供可在组织中诊断入侵细胞水平的方法，包括在CD38BP和潜在的含CD38的组织间形成免疫复合物，然后检测免疫复合物的形成，其中免疫复合物的形成与组织中存在入侵细胞相关。可用标记的分离抗体和标准成像技术在体内进行接触，或可在组织样品上进行体外接触。More specifically, the invention provides for the identification and diagnosis of invading cells and tissues and other cells to which the CD38BPs of the invention are located, thereby monitoring treatment progress, post-treatment status, risk of developing cancer, cancer progression, and the like. In one embodiment of such a diagnostic test, the invention provides a method for diagnosing the level of invading cells in a tissue, comprising forming an immune complex between CD38BP and potential CD38-containing tissue, and then detecting the formation of the immune complex, wherein The formation of immune complexes is associated with the presence of invading cells in the tissue. Contacting can be performed in vivo using labeled isolated antibodies and standard imaging techniques, or contacting can be performed in vitro on tissue samples.

CD38BPs可用于在任何合适的生物样品中提供任何合适的接受检测含CD38的肽和肽片段。本发明提供的传统免疫检测的实施例包括但不限定于，用CD38BP进行ELISA、RIA、FACS检测、表面等离子共 振检测、色谱检测、组织免疫组织化学、Western印迹和/或免疫沉淀。本发明的抗CD38抗体可用于在人中检测CD38和CD38片段。在这些技术中所用的合适的CD38BP和/或二抗的标记包括但不限定于，各种酶、辅基、荧光材料和放射性材料。合适的酶的实施例包括辣根过氧化物酶、碱性磷酸酶β-半乳糖苷酶或乙酰胆碱酯酶；合适的辅基复合物的实施例包括链亲和素/生物素和亲和素/生物素；合适的荧光材料的实施例包括伞形酮、荧光素、异硫氰酸荧光素、若丹明、二氯三嗪基胺、荧光素、丹磺酰氯或藻红蛋白；化学发光材料的实施例包括鲁米诺；合适的放射性材料的实施例包括¹²⁵I、¹³¹I、³⁵S和³H。CD38BPs can be used to provide any suitable detectable CD38-containing peptides and peptide fragments in any suitable biological sample. Examples of traditional immunoassays provided by the present invention include, but are not limited to, ELISA, RIA, FACS detection, surface plasmon resonance detection, chromatographic detection, tissue immunohistochemistry, Western blotting and/or immunoprecipitation using CD38BP. The anti-CD38 antibodies of the invention can be used to detect CD38 and CD38 fragments in humans. Suitable labels for CD38BP and/or secondary antibodies used in these techniques include, but are not limited to, various enzymes, prosthetic groups, fluorescent materials, and radioactive materials. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase β-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/ Biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine, fluorescein, dansyl chloride, or phycoerythrin; chemiluminescent materials Examples of suitable radioactive materials include luminol; examples of suitable radioactive materials include ¹²⁵ I, ¹³¹ I, ³⁵ S, and ³ H.

也可在生物样品中通过竞争免疫检测法，用标记可检测底物的CD38肽标准品和未标记的CD38BP，例如未标记的抗CD38抗体来检测CD38BPs。在这种检测中，生物样品、标记的CD38肽标准品和CD38BP结合，从而测定与未标记CD38BP结合的标记的CD38标准品的量。生物样品中的CD38肽的量与结合到CD38BP上的标记的CD38标准品的量成反比。CD38BPs can also be detected in biological samples by competition immunoassays using a CD38 peptide standard labeled with a detectable substrate and unlabeled CD38BP, eg, an unlabeled anti-CD38 antibody. In this assay, a biological sample, a labeled CD38 peptide standard, and CD38BP are combined to determine the amount of labeled CD38 standard bound to unlabeled CD38BP. The amount of CD38 peptide in the biological sample is inversely proportional to the amount of labeled CD38 standard bound to CD38BP.

CD38BPs在体内肿瘤成像中是特别有用的。与CD38相关的体内成像可通过任何合适技术进行。例如在肿瘤中使用Tc-标记或用其它释放γ射线的同位素标记抗CD38抗体或次级标记(例如FITC标记)来自肿瘤的CD38BP:CD38复合物，并用γ闪烁照相机(例如Elscint Apex409ECT设备)进行成像，典型地使用低能、高分辨率瞄准仪或低能全目标瞄准仪。然后评估染色组织的放射性计数，作为肿瘤中CD38相关肽的量的指示。通过这种技术获得的图像可用于评估CD38在患者、哺乳动物或组织中的生物分布，例如，以CD38或CD38片段作为存在入侵的癌细胞的生物标志物。这种技术的改变可包括使用磁共振成像(MRI)改善通γ照相机的成像技术。类似的免疫成像方法和原理在例如Srivastava(ed.)，Radiolabeled Monoclonal Antibodies For Imaging AndTherapy(Plenum Press 1988)，Chase，″Medical Applications ofRadioisotopes，″in Remington′s PharmaceuticalSciences，18th Edition，Gennaro et al.，(eds.)、pp.624-652(Mack Publishing Co.，1990)，andBrown，″Clinical Use of Monoclonal Antibodies，″in BiotechnologyAndPharmacy 227-49，Pezzuto et al.，(eds.)(Chapman&Hall 1993)中描述。这种图像也可用于其它抗癌试剂的定向递送，其实施例如此处所述 (例如，凋亡试剂、毒素或CHOP化疗组合物)。另外，这种图像也可或选择性地作为外科技术切除肿瘤的依据。另外，这种体内成像技术也可在具有肿瘤的患者中鉴定和定位肿瘤(由于存在其它生物标志物、转移等)，但肿瘤不能通过传统分析技术鉴定。所有这些方法均是本发明的特征。CD38BPs are particularly useful in in vivo tumor imaging. In vivo imaging associated with CD38 can be performed by any suitable technique. For example in tumors use Tc-label or label anti-CD38 antibodies with other gamma-releasing isotopes or secondary label (eg FITC label) CD38BP:CD38 complexes from tumors and image with gamma scintillation camera (eg Elscint Apex409ECT device) , typically using a low-energy, high-resolution collimator or a low-energy all-target collimator. Radioactive counts of the stained tissue were then assessed as an indication of the amount of CD38-related peptide in the tumor. Images obtained by this technique can be used to assess the biodistribution of CD38 in patients, mammals or tissues, for example, using CD38 or CD38 fragments as a biomarker for the presence of invasive cancer cells. Variations of this technique may include the use of Magnetic Resonance Imaging (MRI) to improve imaging techniques with gamma cameras. Similar immunoimaging methods and principles are described, for example, in Srivastava (ed.), Radiolabeled Monoclonal Antibodies For Imaging And Therapy (Plenum Press 1988), Chase, "Medical Applications of Radioisotopes," in Remington's Pharmaceutical Sciences, 18th Edition, Gennaro et al., ( eds.), pp.624-652 (Mack Publishing Co., 1990), and Brown, "Clinical Use of Monoclonal Antibodies," in Biotechnology And Pharmacy 227-49, Pezzuto et al., (eds.) (Chapman & Hall 1993). Such images can also be used for targeted delivery of other anti-cancer agents, examples of which are described herein (e.g., apoptotic agents, toxins, or CHOP chemotherapeutic compositions). In addition, such images may also or alternatively serve as a basis for surgical techniques to remove tumors. Additionally, this in vivo imaging technique can also identify and localize tumors in patients with tumors (due to the presence of other biomarkers, metastasis, etc.), but tumors cannot be identified by traditional analytical techniques. All of these methods are features of the present invention.

本发明提供的体内成像和其它诊断技术在人患者(例如，以前未诊断具有癌症的患者，或在癌症恢复/康复期的患者)中检测微转移是非常有用的。例如，在所有癌细胞中占90％的癌细胞可用CD38抗体偶联组合物染色鉴定。用单克隆抗CD38抗体和其它此处所述的CD38BPs的检测可作为存在侵略性/入侵性癌的暗示，也或选择性地用针对这种微转移的相关单克隆抗CD38抗体、CD38BP或相关组合物治疗来提供可行性的暗示。另外，与癌细胞相关的单克隆抗CD38抗体利于区分这种癌相关组织和来自正常细胞的其它形式CD38相关的细胞。The in vivo imaging and other diagnostic techniques provided by the present invention are very useful in detecting micrometastases in human patients (eg, patients not previously diagnosed with cancer, or patients in recovery/convalescence from cancer). For example, cancer cells accounting for 90% of all cancer cells can be identified by staining with CD38 antibody conjugate composition. Detection with monoclonal anti-CD38 antibodies and other CD38BPs described here may be indicative of the presence of aggressive/invasive cancer, or alternatively with related monoclonal anti-CD38 antibodies, CD38BPs or related Composition treatments to provide a hint of feasibility. In addition, monoclonal anti-CD38 antibodies associated with cancer cells facilitate the distinction of such cancer-associated tissues from other forms of CD38-associated cells from normal cells.

在一个技术方案中，本发明提供体内成像方法，其中本发明的CD38BP，譬如抗CD38抗体与促进检测的无线电不透明试剂偶联，将偶联的抗体给药宿主，譬如通过注入血液中，然后检测宿主中标记抗体的存在和定位。通过这种技术和任何其它此处提供的诊断方法，本发明提供了在人患者或采自人患者的生物样品中筛选疾病相关的细胞存在的方法。对诊断成像而言，可将放射性同位素与CD38BP通过中间功能基团直接或间接结合。有用的中间功能基团包括螯合剂，例如乙二胺四乙酸和二乙烯三胺五乙酸(参见例如US5,057,313)。在这种放射性同位素偶联的CD38BPs参与的诊断检测中，递送到患者的偶联的肽的剂量典型地维持在尽可能低的水平，使同位素具有最小半衰期、在机体具有最小滞留时间，同位素数量最小，从而进行检测和精确测定。In one embodiment, the invention provides a method for in vivo imaging wherein a CD38BP of the invention, such as an anti-CD38 antibody, is conjugated to a radio-opaque reagent that facilitates detection, the conjugated antibody is administered to a host, such as by infusion into the blood, and detection is then performed. Presence and localization of labeled antibodies in the host. By this technique and any other diagnostic method provided herein, the present invention provides methods of screening for the presence of disease-associated cells in a human patient or a biological sample taken from a human patient. For diagnostic imaging, radioactive isotopes can be directly or indirectly combined with CD38BP through intermediate functional groups. Useful intermediate functional groups include chelating agents such as ethylenediaminetetraacetic acid and diethylenetriaminepentaacetic acid (see eg US 5,057,313). In diagnostic assays involving such radioisotope-conjugated CD38BPs, the dose of the conjugated peptide delivered to the patient is typically kept as low as possible to minimize the half-life of the isotope, the minimum residence time in the body, the amount of isotope Minimal for detection and precise measurement.

除了放射性同位素和无线电不透明试剂外，可使用与染料(譬如偶联生物素-链亲和素复合物)、造影剂、荧光化合物或分子及用于磁共振成像(MRI)的增强试剂(例如顺磁性离子)(参见例如美国专利6,331,175，它描述了MRI技术和与MRI增强试剂偶联的抗体的制备)偶联的CD38BPs进行诊断方法。这种诊断/检测试剂可选自用于磁共振成像的试剂和荧光化合物。为了加载诸如抗体，偶联放射性金属或顺磁性离子的成分这样的CD38BP，需要使其与具有连接多个可结合离子的螯合剂基团的长尾的试剂反应。这种尾可以是多聚体，譬如多聚赖氨酸、多糖 或其它衍生的或具有可与螯合剂基团，譬如例如卟啉、多胺、冠醚、二缩氨基硫脲、聚肟等已知用于该目的的悬挂基团的衍生链。可用标准化学法将螯合剂与CD38BPs偶联。螯合剂通常通过可与分子形成具有最小免疫反应性丢失和最小聚集和/或内部交联的基团与CD38BP连接。其它不常用的将螯合剂与抗体偶联的方法和试剂在例如US4,824,659中描述。潜在有用的金属螯合剂组合物的实施例包括2-苯甲基-DTPA及其一甲基和环己基类似物，使用一般能量范围为60到4,000keV的诊断性同位素，例如¹²⁵I、¹²³I、¹²⁴I、⁶²Cu、⁶⁴Cu、¹⁸F、¹¹¹In、⁶⁷Ga、⁶⁷Ga、⁹⁹Tc、 ⁹⁴Tc、¹¹C、¹³N、¹⁵O和⁷⁶BR进行放射性成像。这些和类似的螯合剂在与非放射性金属，譬如镁、铁和钆结合时，可与CD38BPs联合用于MRI诊断方法。大环螯合剂，例如NOTA、DOTA和TETA可与各种金属和放射性金属一起使用，特别是分别与镓、钇和铜的放射性核一起使用。可通过剪辑目标金属的环体积而使这种金属-螯合剂复合物非常稳定。其它诸如大环聚醚这样的与核素，例如进行RAIT的²²³Ra稳定结合的环状螯合剂也适用于诊断方法中。In addition to radioisotopes and radio-opaque agents, dyes (such as conjugated biotin-streptavidin complexes), contrast agents, fluorescent compounds or molecules, and enhancing reagents for magnetic resonance imaging (MRI) (such as cis Magnetic ions) (see, eg, US Pat. No. 6,331,175, which describes MRI techniques and preparation of antibodies conjugated to MRI-enhancing reagents) conjugated CD38BPs for diagnostic methods. Such diagnostic/detection reagents may be selected from reagents and fluorescent compounds used in magnetic resonance imaging. To load a CD38BP such as an antibody, a component conjugated to a radioactive metal or a paramagnetic ion, it needs to be reacted with a reagent having a long tail linking multiple ion-binding chelator groups. Such tails can be polymeric, such as polylysine, polysaccharide or other derivatized or have chelating agent groups, such as for example porphyrin, polyamine, crown ether, thiosemicarbazone, polyoxime, etc. Derivatized chains of pendant groups are known for this purpose. Chelators can be coupled to CD38BPs using standard chemistry. Chelators are typically attached to CD38BP through groups that can form with the molecule with minimal loss of immunoreactivity and minimal aggregation and/or internal crosslinking. Other less common methods and reagents for coupling chelators to antibodies are described, for example, in US 4,824,659. Examples of potentially useful metal chelator compositions include 2-benzyl-DTPA and its monomethyl and cyclohexyl analogs, using diagnostic isotopes such as ¹²⁵ I, ¹²³ I , ¹²⁴ I, ⁶² Cu, ⁶⁴ Cu, ¹⁸ F, ¹¹¹ In, ⁶⁷ Ga, ⁶⁷ Ga, ⁹⁹ Tc, ⁹⁴ Tc, ¹¹ C, ¹³ N, ¹⁵ O and ⁷⁶ BR for radiographic imaging. These and similar chelators, when bound to non-radioactive metals such as magnesium, iron and gadolinium, can be used in MRI diagnostic methods in conjunction with CD38BPs. Macrocyclic chelators such as NOTA, DOTA and TETA can be used with various metals and radiometals, especially with radionuclides of gallium, yttrium and copper, respectively. This metal-chelator complex can be made very stable by clipping the ring volume of the target metal. Other cyclic chelators such as macrocyclic polyethers that stably bind nuclides such ^as223Ra to perform RAIT are also suitable for use in diagnostic methods.

因此，本发明提供诊断型CD38BP偶联物，其中CD38BP与造影剂(例如用于磁共振成像、计算机X线断层摄影术或超声造影剂增强试剂)或放射性核素，例如γ-、β-、α-、俄歇电子或正电子发射同位素偶联。其它有用的偶联CD38BPs在其它地方描述，可用于本发明提供的诊断方法和组合物(例如诊断试剂盒)中。Accordingly, the present invention provides diagnostic CD38BP conjugates, wherein CD38BP is combined with a contrast agent (e.g. for magnetic resonance imaging, computed tomography or ultrasound contrast agent enhancing reagents) or a radionuclide, such as γ-, β-, α-, Auger electron or positron emitting isotope coupling. Other useful conjugated CD38BPs are described elsewhere and can be used in the diagnostic methods and compositions (eg, diagnostic kits) provided herein.

在一个技术方案中，本发明提供诊断癌症的试剂盒，包括含诸如抗CD38抗体这样的CD38BP和一种或多种检测CD38BP与CD38肽结合的试剂的试剂盒。例如，试剂可包括荧光标签、酶标签或其它可检测标签。试剂也可包括二抗和三抗及用于酶反应的试剂，其中酶反应可产生可显现的产物。在一个技术方案中，本发明提供诊断试剂盒，在合适的容器中含一种或多种标记或未标记形式的本发明诸如抗CD38抗体这样的CD38BPs，间接检测法中的温育试剂，及根据标记的状态，在这种检测中进行检测的底物或衍生的试剂。也包括对照试剂和用途说明书。In one technical solution, the present invention provides a kit for diagnosing cancer, including a kit comprising CD38BP such as an anti-CD38 antibody and one or more reagents for detecting the combination of CD38BP and CD38 peptide. For example, reagents may include fluorescent tags, enzyme tags, or other detectable tags. Reagents may also include secondary and tertiary antibodies and reagents for enzymatic reactions that result in visualized products. In one embodiment, the invention provides a diagnostic kit comprising, in a suitable container, one or more CD38BPs of the invention, such as an anti-CD38 antibody, in labeled or unlabeled form, an incubation reagent in an indirect assay, and Depending on the state of labelling, the substrates or derivatized reagents are detected in this assay. Control reagents and instructions for use are also included.

也提供了诊断型试剂盒，通过诸如偶联/标记的抗CD38抗体这样的CD38BP在组织样品或宿主中检测细胞活性或检测CD38肽的存在。在这种诊断型试剂盒及其它地方所述的用于治疗用途的试剂盒中，典型地可在试剂盒中以冻干形式，单独或与其它针对靶细胞或肽的抗体一起提 供CD38BP。典型地，也包括药学上可接受载体(例如惰性稀释剂)和/或其成分，例如Tris、磷酸或碳酸缓冲液、稳定剂、防腐剂、生物杀灭剂、生物杀灭剂、惰性蛋白，例如血清白蛋白等(典型地，装在单独容器中，用于混合)及其它试剂(典型地也在单独的容器中)。在特定试剂盒中，也包括可结合抗CD38抗体或其它CD38BP的第二抗体，它典型地存在单独的容器中。第二抗体典型地以与本发明的抗CD38抗体或其它CD38BP相似的方式与标记物偶联和配制。用上述方法CD38BPs可用于区分癌/肿瘤细胞的亚组，并鉴定这种细胞及相关的组织/生长。Also provided are diagnostic kits for detecting cellular viability or detecting the presence of CD38 peptides in tissue samples or hosts via CD38BP such as conjugated/labeled anti-CD38 antibodies. In such diagnostic kits, as well as kits for therapeutic use as described elsewhere, CD38BP will typically be provided in the kit in lyophilized form, alone or with other antibodies directed against the target cell or peptide. Typically, pharmaceutically acceptable carriers (such as inert diluents) and/or components thereof, such as Tris, phosphate or carbonate buffers, stabilizers, preservatives, biocides, biocides, inert proteins, are also included, Examples include serum albumin etc. (typically in separate containers for mixing) and other reagents (also typically in separate containers). In certain kits, a secondary antibody that binds the anti-CD38 antibody or other CD38BP is also included, typically in a separate container. Secondary antibodies are typically conjugated and formulated to the marker in a similar manner to the anti-CD38 antibodies or other CD38BPs of the invention. Using the methods described above CD38BPs can be used to differentiate subgroups of cancer/tumor cells and to identify such cells and associated tissues/growths.

在一个实施例中，CD38BP或抗CD38抗体可加到硝酸纤维素或其它可固定细胞、细胞颗粒或可溶性蛋白的固相支持物上。然后在用可检测标记的CD38肽或抗体处理后用合适的缓冲液洗涤支持物。然后用缓冲液洗涤固相支持物以除去未结合的肽或抗体。然后可通过已知的方法步骤检测固相支持物上结合的标记物的量。In one embodiment, CD38BP or an anti-CD38 antibody can be added to nitrocellulose or other solid supports that can immobilize cells, cell particles or soluble proteins. The support is then washed with a suitable buffer after treatment with the detectably labeled CD38 peptide or antibody. The solid support is then washed with buffer to remove unbound peptide or antibody. The amount of bound label on the solid support can then be detected by known method steps.

连接的与暴露的底物反应的酶可用于产生可被检测的化学基团，例如，通过分光光度测定、荧光测定或视觉观察的方法检测CD3 8BP偶联物和/或融合蛋白。可用作可检测标记CD38BPs和抗CD38抗体的酶包括苹果酸脱氢酶、葡萄球菌核酸酶、δ-5-类固醇异构酶、酵母乙醇脱氢酶、α-甘油磷酸脱氢酶、丙糖磷酸异构酶、辣根过氧化物酶、碱性磷酸酶、天冬酰胺酶、葡萄糖氧化酶、β-半乳糖苷酶、核糖核酸酶、尿素酶、过氧化氢酶、葡萄糖-6-磷酸脱氢酶、葡糖淀粉酶和乙酰胆碱酯酶。也可以用荧光化合物标记CD38BP。当荧光标记的抗体暴露在合适波长的光下时，可通过荧光检测其存在。最常用的荧光标记化合物是荧光素异硫氰酸盐、若丹明、藻红蛋白、藻蓝蛋白、异藻蓝蛋白、o-邻苯二甲醛和荧光胺。Linked enzymes that react with exposed substrates can be used to generate chemical moieties that can be detected, for example, by spectrophotometric, fluorometric, or visual inspection of CD3 8BP conjugates and/or fusion proteins. Enzymes that can be used as detectable markers for CD38BPs and anti-CD38 antibodies include malate dehydrogenase, staphylococcal nuclease, delta-5-steroid isomerase, yeast alcohol dehydrogenase, alpha-glycerophosphate dehydrogenase, triose Phosphate isomerase, horseradish peroxidase, alkaline phosphatase, asparaginase, glucose oxidase, beta-galactosidase, ribonuclease, urease, catalase, glucose-6-phosphate Dehydrogenase, glucoamylase, and acetylcholinesterase. CD38BP can also be labeled with a fluorescent compound. The presence of fluorescently labeled antibodies can be detected by fluorescence when exposed to light of the appropriate wavelength. The most commonly used fluorescently labeled compounds are fluorescein isothiocyanate, rhodamine, phycoerythrin, phycocyanin, isophycocyanin, o-phthalaldehyde, and fluorescamine.

诸如抗CD38抗体的CD38BPs也可用荧光发射的金属，譬如¹⁵²Eu或其它镧系列金属进行可检测标记。例如，这些金属可被抗CD38抗体通过金属螯合基团例如二乙三胺五乙酸(DTPA)或乙二胺四乙酸(EDTA)进行连接。CD38BPs such as anti-CD38 antibodies can also be detectably labeled with fluorescent emitting metals such ^as152Eu or other lanthanum series metals. For example, these metals can be attached by anti-CD38 antibodies via metal chelating groups such as diethylenetriaminepentaacetic acid (DTPA) or ethylenediaminetetraacetic acid (EDTA).

CD38BPs和抗CD38抗体也可通过与化学发光化合物偶联而进行可检测标记。然后通过检测在化学反应中升高的荧光的存在来测定化学发光标记的CD38-BP的存在。特别有用的化学发光标记化合物的实施例有发光氨、异氨基苯二酰肼、theromatic吖啶酯、咪唑、吖啶盐和草酸酯。CD38BPs and anti-CD38 antibodies can also be detectably labeled by conjugation with chemiluminescent compounds. The presence of chemiluminescence-labeled CD38-BP is then determined by detecting the presence of fluorescence that increases during the chemical reaction. Examples of particularly useful chemiluminescent labeling compounds are luminol, isoaminophthalhydrazide, theromatic acridinium esters, imidazoles, acridinium salts and oxalates.

同样，生物发光化合物可用于标记CD38BP。生物发光是一种在生物系统中发现的化学发光，其中催化蛋白可增加化学发光反应的效率。生物发光蛋白的存在通过检测荧光而测定。用于标记的重要的生物发光化合物有荧光素、荧光素酶和发光蛋白质。例如如果可检测标记是放射性γ发射者，则标记肽或抗体、抗体片段或衍生物的检测可通过闪烁计数来完成，如果标记是荧光材料，则通过荧光计数来检测。对酶标记而言，可通过使用酶底物的比色方法进行检测。也可通过将底物的酶反应程度与类似制备的标准品进行视觉比较而进行检测。Likewise, bioluminescent compounds can be used to label CD38BP. Bioluminescence is a type of chemiluminescence found in biological systems where catalytic proteins increase the efficiency of the chemiluminescent reaction. The presence of bioluminescent proteins is determined by detecting fluorescence. Important bioluminescent compounds for labeling are luciferin, luciferase and luminescent proteins. For example, detection of the labeled peptide or antibody, antibody fragment or derivative can be accomplished by scintillation counting if the detectable label is a radioactive gamma emitter, or by fluorescence counting if the label is a fluorescent material. For enzyme labels, detection can be performed by colorimetric methods using enzyme substrates. It can also be detected by visual comparison of the extent of enzymatic reaction of the substrate with similarly prepared standards.

这些和其它诊断技术可用于筛选任何合适的针对CD38肽或CD38片段的材料。这些被筛选的材料的实施例包括例如血液、血清、淋巴、尿液、炎症流出液、脑脊液、羊水、组织提取物或组织匀浆等。但本发明不限定仅检测这些样品，本领域技术人员可确定使用其它样品的合适条件。These and other diagnostic techniques can be used to screen any suitable material against CD38 peptides or CD38 fragments. Examples of these materials to be screened include, for example, blood, serum, lymph, urine, inflammatory effluent, cerebrospinal fluid, amniotic fluid, tissue extracts or tissue homogenates, and the like. However, the present invention is not limited to testing only these samples, and those skilled in the art can determine the appropriate conditions for using other samples.

可通过从患者中除去组织样品，并对这种样品提供本发明的标记CD38BPs，譬如抗CD38抗体而进行原位检测。通过对生物样品使用或覆盖标记的CD38BP，譬如标记的抗CD38抗体(或片段)，可提供本发明的CD38BP、抗CD38抗体(或片段)。通过使用这种流程，不仅可测定CD38或CD38片段的存在，也可以测定这种肽在被检测的组织中的分布(例如，以评估癌细胞的散布)。通过本发明，技术人员应知道，可改变任何各种组织学方法(例如染色流程)以进行原位检测。In situ detection can be performed by removing a tissue sample from a patient and providing this sample with labeled CD38BPs of the invention, such as an anti-CD38 antibody. The CD38BP, anti-CD38 antibody (or fragment) of the present invention can be provided by using or covering a labeled CD38BP, such as a labeled anti-CD38 antibody (or fragment), on a biological sample. By using this protocol, not only the presence of CD38 or CD38 fragments can be determined, but also the distribution of this peptide in the tissue being examined (eg, to assess the dissemination of cancer cells). With the present invention, the skilled artisan will appreciate that any of the various histological methods (eg, staining protocols) can be modified for in situ detection.

本发明进一步提供促进销售和/或使用本发明的CD38BP的方法，包括发布关于对任何可能的人或实体在预防或治疗任何如其它地方所述的疾病或疾病组合方面化合物的用途信息(例如提供分发、邮寄等的印刷资料，通过广告标记，通过电视节目和广告，通过收音机节目和广告，通过互联网邮寄、通过电子邮件、通过电话销售、通过门对门或人对人销售、通过基金和/或集会、座谈、论坛等、通过使用和/或缩减销售人员的服务和/或医疗/科学联络、通过基金和/或集合科学研究及相关用途的出版物等)(譬如，药物链、规定的管理者、保险公司、HMOs、医院和医院链、其它康复公司、药房管理者、潜在患者、癌症患者、以前的癌症患者、恢复期患者、初级保健医生、护士、药房医生和/或关键的意见领导者)。The invention further provides methods of promoting the sale and/or use of the CD38BPs of the invention, including disseminating information about the use of the compound to any possible person or entity in the prevention or treatment of any disease or combination of diseases as described elsewhere (e.g., providing Printed materials distributed, mailed, etc., by advertising labels, by television programs and commercials, by radio programs and commercials, by Internet mail, by electronic mail, by telephone sales, by door-to-door or person-to-person sales, by funds and/or Meetings, symposiums, forums, etc., by using and/or reducing the services of sales staff and/or medical/scientific liaison, by funding and/or gathering scientific research and publications for related purposes, etc.) (e.g., pharmaceutical chains, prescribed regulatory providers, insurance companies, HMOs, hospitals and hospital chains, other rehab companies, pharmacy managers, prospective patients, cancer patients, former cancer patients, recovering patients, primary care physicians, nurses, pharmacy physicians, and/or key opinion leaders By).

本发明也提供含本发明的化合物的药物组合物及使用说明书。试剂 盒可进一步含有一种或多种其它试剂，例如上述的免疫抑制试剂、化疗试剂和抗炎症试剂或放射性毒素试剂，或一种或多种本发明的其它CD38BPs(例如具有补体活性的CD38BP)。本发明的试剂盒也包括诊断试剂和/或其它治疗试剂。在一个技术方案中，本发明的试剂盒包括用于诊断方法中以诊断患者中表达CD38的细胞参与的疾病状态或存在的本发明的CD38BP和诊断试剂。在一个技术方案中，试剂盒包括以高稳定形式(例如冻干形式)存在的本发明的CD38BP，它可与能和高稳定组合物混合形成可注射组合物的药学上可接受的载体一起存在。The invention also provides pharmaceutical compositions comprising the compounds of the invention and instructions for their use. The kit may further contain one or more other reagents, such as the above-mentioned immunosuppressive reagents, chemotherapeutic reagents and anti-inflammatory reagents or radiotoxic reagents, or one or more other CD38BPs of the present invention (such as CD38BPs with complement activity) . The kits of the invention also include diagnostic and/or other therapeutic reagents. In one technical solution, the kit of the present invention comprises the CD38BP of the present invention and a diagnostic reagent for use in a diagnostic method to diagnose a disease state in which CD38-expressing cells are involved or present in a patient. In one technical solution, the kit includes the CD38BP of the present invention in a highly stable form (such as a lyophilized form), which can be present together with a pharmaceutically acceptable carrier that can be mixed with a highly stable composition to form an injectable composition .

此处引用的所有参考文献，包括出版物、专利申请和专利均通过与每个参考文献单独和特异地通过参考整合在文中的程度进行参考并整合在文中。All references, including publications, patent applications, and patents, cited herein are referenced and incorporated herein to the extent that each reference is individually and specifically incorporated by reference.

此处所用的所有标题和副标题仅为了方便，不应对本发明以任何方式进行限制。All headings and subheadings used herein are for convenience only and shall not limit the invention in any way.

上述元素在其所有可能变化下的任何组合都包括在本发明中，除非进行说明或与本文明确矛盾。Any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

术语“a”和“an”和“the”及类似指示物在描述本发明的文中解释为包括单数和复数，除非进行说明或与本文明确矛盾。The terms "a" and "an" and "the" and similar referents in the context of describing the invention are to be construed as including the singular and the plural unless stated otherwise or clearly contradicted by the context.

此处的数值引用范围仅用于作为落在该范围中的每个单独数值的速记方法，除非特别说明，每个单个的数值包括在说明书中，和它是单独引用一样。除非特别说明，所有此处提供的确切数值是相应近似数值的代表(例如，对特定因素或测定法提供所有确切的示例数值可认为也提供了相应的近似测定值，当合适时用“约”修饰)。Recitation of ranges of values herein are intended merely as a shorthand method of referring to each individual value falling within the range, and unless otherwise indicated, each individual value is included in the specification as if it were individually recited. Unless otherwise indicated, all exact numerical values provided herein are representative of corresponding approximate numerical values (for example, providing all exact exemplary values for a particular factor or assay is considered to also provide the corresponding approximate measurement value, and the use of "about" when appropriate modified).

此处所述所有方法可以合适的顺序进行，除非特别说明或在文中明确矛盾。All methods described herein can be performed in a suitable order unless otherwise indicated herein or otherwise clearly contradicted by context.

此处提供的任何和全部实施例或示例语言(例如“譬如”)的用途仅仅是为了更好地阐述本发明，不对本发明的范围进行限制，除非说明。说明书中的语言不表明任何元素对本发明的实施是必需的，除非明确说明。The use of any and all examples, or exemplary language (eg, "such as") provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise stated. No language in the specification indicates that any element is essential to the practice of the invention unless explicitly stated otherwise.

此处的专利文献的引用和整合仅为了方便，不代表任何该专利文献关于有效性、专利权和/或主张。The citation and integration of the patent documents herein is for convenience only, and does not represent any validity, patent rights and/or claims of the patent documents.

在本发明的任何技术方案的描述中所用术语例如“含有”、“具有”、“包括”或“包含”某成分是指对本发明类似技术方案中“由......组成”、 “主要由......组成”或“主要含有”该特定成分提供支持，除非特别说明或在文中明显有矛盾(例如，此处含特定成分的组合物也理解为描述由该成分组成的组合物，除非特别说明或在文中有明显矛盾)。在法律允许的最大程度上，本发明包括所有技术方案中再次引用的主体事件的修改和等价物。Terms used in the description of any technical solution of the present invention such as "comprising", "having", "comprising" or "including" a certain component refer to "consisting of", "consisting of" in similar technical solutions of the present invention Consisting essentially of" or "consisting essentially of" the specified ingredient, unless specifically stated or otherwise clearly contradicted in the context (for example, a composition containing a particular ingredient herein is also understood to describe a composition consisting of that ingredient. Compositions, unless otherwise specified or clearly contradicted by the text). To the maximum extent permitted by law, the present invention includes modifications and equivalents of the subject matters recited in all technical solutions.

所有此处引用的专利、正在审理的专利申请和其它文献通过参考整合在文中。All patents, pending patent applications, and other documents cited herein are incorporated by reference.

通过以下实施例进一步描述本发明，这些实施例不进行进一步限制。The invention is further described by the following examples, which are not further limited.